Research ArticleEvaluation of the Antioxidative Antibacterial andAnti-Inflammatory Effects of the Aloe FermentationSupernatant Containing Lactobacillus plantarum HM2187491

Meixiu Jiang1 Kan Deng1 Chunling Jiang2 Mingui Fu3 Chunlan Guo4 Xiaolei Wang1

Xin Wang1 Fanjing Meng1 Shaoguo Yang1 Keyu Deng1 Tingtao Chen1 and Hongbo Xin1

1 Institute of Translational Medicine Nanchang University Nanchang Jiangxi 330031 China2Jiangxi Cancer Hospital Nanchang 330045 China3Department of Basic Medical Science School of Medicine University of Missouri Kansas City Kansas City MO 64108 USA4College of Forestry Jiangxi Agriculture University Nanchang 330045 China

Correspondence should be addressed to Tingtao Chen chentingtao1984163com and Hongbo Xin hongboxinyahoocom

Received 12 March 2016 Revised 17 May 2016 Accepted 30 May 2016

Academic Editor Teresa Zelante

Copyright copy 2016 Meixiu Jiang et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Little work is done to develop Aloe vera (AV) using probiotics To explore the potential benefits the antioxidant effects and theantibacterial effects on foodborne pathogens of Aloe fermentation supernatant were evaluated in vitro Our results indicated thatthe Aloe fermentation supernatant fermented by Lactobacillus plantarum HM2187491 had very strong scavenging capacities ofthe DPPH (86) O

2

∙minus (85) ∙OH (76) and Fe2+ chelation (82) and reducing powers (2425mgL) and the inhibition zonesfor Salmonella typhimurium Salmonella enteritidis Shigella flexneri Escherichia coli Listeria monocytogenes S dysenteriae 301Staphylococcus aureusCowan1 and Propionibacterium acneswere 16 15 19 20 21 20 and 27mmMoreover the low concentrationof Aloe fermentation supernatant had significantly reduced the production of IL-1120573 TNF-120572 and IL-6 in both mRNA and proteinlevels (119875 lt 001)Therefore theAloe fermentation supernatant can be used as functional beverage or cosmetic ingredients to guardhuman intestinal health delaying senescence and prevent chronic diseases

1 Introduction

Aloe vera (AV) is a cactus-like plant which has been usedwidely in herbal medicines for millennia [1]The compoundsin fleshy leaves for example aloe emodin aloin aloesinsaponins terpenoids and polysaccharides possess thewound healing anti-inflammatory immunity antidiabeticantioxidant laxative antibacterial antifungal antiviral andantitumor effects [2ndash6] Now AVhas also beenwidely used ascosmetic-moisturizers tooth pastes food flavoring com-pounds and preservative in the pharmaceutical and foodfields [2] However little work is done to exploit the AV usingprobiotics

Probiotics are defined as ldquolive microorganisms whichwhen administered in adequate amounts confer a health ben-efit on the hostrdquo Previous studies indicated that probiotics

conferred miscellaneous health benefits include an improve-ment in lactose intolerance an increase in natural resistanceto infectious disease in the gastrointestinal tract the suppres-sion of cancer an antidiabetic agent and a decrease in serumcholesterol levels [7ndash9] In recent decades probiotics are thehotspot for microbiologist and nutritionists [10 11] and thehealth awareness and busy lifestyle have created a strong anddynamic probiotics market which offers good prospects forconsumers [12]

Among the probiotics Lactobacillus plantarum is pointedout as an industrially important microorganism that can befound and isolated from dairy products fermented foods(sauerkraut sourdough sausages cheeses wines olives andpickled vegetables) environments (cow-dung silage andsewage) human mouth intestinal tract and stools [13 14]and this strain had been widely used for the development of

Hindawi Publishing CorporationMediators of InflammationVolume 2016 Article ID 2945650 8 pageshttpdxdoiorg10115520162945650

2 Mediators of Inflammation

functional foods and potential oral vaccines [13] In ourprevious work [15] L plantarum HM2187491 was isolatedfrom human intestinal which could grow better at 37∘C in ananaerobic environment and could resist harsh conditions ofgastrointestinal tract (pH 25 and 030 bile salt) Moreoverthis strain could strongly adhere toHT-29 cells and reduce theadhesion of Shigella dysenteriae 2457 Staphylococcus aureusCowan1 Enterobacter sakazakii 45401 and Escherichia coli44102 to HT-29 cells and its long-term residence in miceintestine increased the number of lactobacilli and decreasedthe number of enterococci [15]

In the present study the tested probiotics (eg Bifi-dobacterium longumATCC15708 L plantarumHM218749 LrhamnosusGG and Streptococcus thermophilusG1) grew wellin AV substrate and conferred AV sound antioxidant andantibacterial effects and the L plantarum HM2187491 wasfinally chosen for its popularity in food fields and perfectbeneficial effects verified in our previous studies and thecell experiment was carried out to further verify the anti-inflammatory effects of the Aloe fermentation supernatant

2 Materials and Methods

21 Preparation of AV Extract and Aloe Fermentation Super-natant TheAV leaves were freshly harvested by cutting fromthe bottom and were washed thoroughly The edges wereremoved and the leaves were peeled to obtain the slimy andtransparent gel contents The gel was then mashed using apulper which was then centrifuged at 4000timesg for 10min toobtain the AV extract [1]

The probiotic Bifidobacterium longum ATCC15708 (iso-lated from the milk) L plantarum HM218749 (isolated fromhuman intestine) L rhamnosusGG (isolated from the milk)and Streptococcus thermophilus G1 (isolated from the milk)were cultured in de Man Rogosa Sharpe (MRS) medium (forLactobacilli and Bifidobacterium) and brainheart infusion(BHI) medium (for Streptococcus thermophilus G1) for 24 hand 108 cfumL of the strains was used as an inoculum to beadded to Aloe vera Aloe vera + 5 glucose or Aloe vera +5 glucose + 5 skimmed milk for preparing the Aloe fer-mentation supernatant which was incubated for 15 to 18 h at37∘C or 42∘C Finally the Aloe fermentation supernatant andAV were freeze-dried using vacuum freeze drier under withthe following conditions freeze drying at 02mbar vacuumminus40∘C for 48 h The lyophilized powder was stored at minus20∘Cfor further use

22 Cultivable Bacterial Counts Bacterial counts were enu-merated as previously reported [16] Diluted aliquots werereplica-plated onto BHI agar and MRS agar and then incu-bated anaerobically in the anaerobic system (full of 85 N

2

5H2 and 10CO

2) at 37∘Cor 42∘C for 24sim36 h followed by

counting colonies from plates showing 25ndash250 colonies [17]

23 Antioxidative and Antibacterial Activity of AV Extract andAloe Fermentation Supernatant For preparation of the Aloefermentation supernatant 10 of the probiotics were addedto AV extract and fermented for 15 to 18 h at 37∘C and thenthe Aloe fermentation cultures were centrifuged at 4000timesg

for 10min and the pellets in the bottom of the centrifugetubes (containing insoluble materials orand probiotics) weredeemed as the residual substance and the clear water insolu-ble substance was deemed as supernatant and the AV extractwas used as a control Then the clearance of the DPPHO2

∙minus ∙OH Fe2+ chelation and oxygen-reduction activitywere measured just as in [18] For antimicrobial activityovernight (12 h) cultures of the pathogenic microorganismsincluding Salmonella typhimurium ATCC 13311 S enteritidisATCC13076 S flexneri ATCC 12022 E coli 44102 Listeriamonocytogenes ATCC 19111 S dysenteriae 301 S aureusCowan1 and Propionibacterium acnes ATCC 11827 werespread on the surface of the LB agar plates and the culturesupernatant (200120583L) was loaded into an Oxford cup (outerdiameter 78 plusmn 01mm inner diameter 60 plusmn 01mm andheight 100plusmn01mm) which was placed on the surface of theagarThe size of the inhibition zone was measured until a for-mation of clear zone around the Oxford cupThe experimentwas carried out in duplicate [17]

24 Cell Viability Assay RAW 2647 cells (passages 8ndash20)were maintained in DMEM supplemented with 10 FBSand antibiotics (100UmL of penicillin and 100 120583gmL ofstreptomycin) at 37∘C in a 5 CO

2incubator Emodin

(Wuhan qi Boster Biological Technology co Ltd) polysac-charide (Wuhan qi Boster Biological Technology co Ltd)Aloe vera (lyophilized powder) and Aloe vera + L plantarum(lyophilized powder) were dissolved in phosphate-bufferedsaline (PBS) solution and their effect on cell viability of RAW2647 cells was determined by MTT assay [19]

RAW 2647 cells were plated at a density of 10000 perwell in 100 120583L of complete culture medium and treatedwith designed concentrations of emodin (0 1 25 5 and10 120583gmL) polysaccharide (0 25 5 15 30 and 50 120583gmL)Aloe vera (0 5 15 30 50 100 and 200120583gmL) and Aloe vera+ L plantarum (0 5 15 30 50 100 and 200120583gmL) in 96-wellmicrotiter plates for 36 h at 37∘C in a humidified incubatorControl cells treated with PBS served as the vehicle groupEach concentration of drugs was repeated in 5 wells Afterincubation for specified times MTT reagent (20120583L 5mgmL) was added to each well and incubated for 4 h After care-ful removal of theMTT solution 150 120583L of DMSOwas addedto each well The absorbance was recorded on a microplatereader at the wavelength of 570 nm

25 Anti-Inflammatory Evaluation of AV Extract and AloeFermentation Supernatant Using RT-PCR and ELISA RAW2647 cells were seeded into 24-well plates (5 times 104 cells perwell) for 24 h and were then washed with PBS Cells weretreatedwithwithout lipopolysaccharide (LPS 1 120583gmL each)emodin (25120583gmL and 10 120583gmL) polysaccharide (25120583gmL and 15 120583gmL) Aloe vera (5 120583gmL and 50 120583gmL) andAloe vera + L plantarum (5 120583gmL and 50120583gmL) for 24 hand then the cells and supernatants were harvested for Q-PCR and ELISA analysis respectively

For the evaluation of cytokine mRNA expression levelstotal RNAs from the RAW 2647 cells were prepared byadding TRIzol reagent (Gibco BRL Grand Island NY USA)according to the manufacturerrsquos protocol and 24 120583g of total

Mediators of Inflammation 3

B longumL plantarum

L rhamnosusS thermophilus

Lg C

FUm

L

30 37 4225

Temperature (∘C)

8

9

10

11

12

(a)

B longum L plantarum L rhamnosus S thermophilus

Lg C

FUm

L

Aloe veraAloe vera + glucoseAloe vera + glucose + skimmed milk

8

9

10

11

12

lowastlowast

lowastlowast lowastlowast lowastlowast lowastlowast

lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast

(b)

Figure 1 The suitable temperature (a) and fermentation substrates (b) for B longum L plantarum L rhamnosus GG and S thermophilusThe significant analysis was done between Aloe vera Aloe vera + glucose and Aloe vera + glucose + skimmed milk Data are shown as themean plusmn SD lowastlowast119875 lt 001

RNA from each group was reverse transcribed to cDNAusing a commercially available kit (Applied Biosystems)Quantitative real-time PCRwas performed with 7900HT fastreal-time PCR system (ABI Foster City CA) using 2 times SYBRGreen master mix (Bio-Rad) Forty cycles were conducted asfollows 95∘C for 30 s and 60∘C for 30 s preceded by 1minat 95∘C for polymerase activation with the following primers(Q-PCR IL-1120573 sense primer 51015840-GTGTCTTTCCCGTGG-ACCTTC1015840-31015840 antisense primer 51015840-TCATCTCGGAGCCTG-TAGTGC-31015840 Q-PCR TNF-120572 sense primer 51015840-GTGGAA-CTGGCAGAAGAGGCA antisense primer 51015840-AGAGGG-AGGCCATTTGGGAAC-31015840 Q-PCR IL-6 sense primer 51015840-GGAAATCGTGGAAATGAG-31015840 antisense primer 51015840-GCT-TAGGCATAACGCACT-31015840 Q-PCR GAPDH sense primer51015840-CTCGTGGAGTCTACTGGTGT-31015840 antisense primer 51015840-GTCATCATACTTGGCAGGTT-31015840)

The products of IL-1120573 TNF-120572 and IL-6 in cell super-natants were determined using the ELISA kit for IL-1120573(eBioscience) TNF-120572 (eBioscience) and IL-6 (eBioscience)

26 Data Analysis Data are reported as means plusmn SD andresults were analyzed using SPSS 130 software (SPSS IncChicago IL USA) by means of independent one-wayANOVA tests in each sampling point The differences amongthe three groups were assessed by means of the least signifi-cant difference (LSD) multiple comparison test (119875 lt 005 or001)

3 Results

31 Exploration of the Fermentation Conditions of Probi-otics To determine the optimum growth temperature of theselected probiotics the same initial inoculums of B longum(MRS medium) L plantarum (MRS medium) L rhamnosusLGG (MRS medium) and S thermophilus (BHI medium)

were inoculated in the corresponding media and cultured at25∘C 30∘C 37∘C and 42∘C and the results showed that theoptimum temperature for B longum L plantarum and Lrhamnosus LGG was 37∘C and S thermophilus received itsmaximum yield at 42∘C (Figure 1(a))

To further investigate the effects of different additives onthe growth of probiotics glucose and skimmed milk wereadded in the AV extract and the results indicated that all thetested strains grew well in AV extract with the initial pH 60(about 9 LgCFUmL Figure 1(b)) It seemed that the additionof 5 glucose and 5 glucose + 5 skimmed milk had sig-nificantly enhanced the biomass of B longum L plantarumand L rhamnosus GG (119886 lt 001) and the growth promotingeffect of the 5 glucose + 5 skimmed milk was muchbetter than 5 glucose (119887 lt 001) (Figure 1(b))

32 Antioxidative and Antibacterial Activity of Aloe Fermen-tation Supernatant To evaluate the additive of the probioticson the perfect antioxidant and antibacterial activities of AV[9 20ndash22] the AV + 5 glucose which gives us more choicesfor the future products was chosen for our next study

Compared with the AV the addition of probiotics madelittle change on O

2

∙minus clearance Fe2+ chelation and reductionactivity while the L plantarumHM2187491 had significantly(119875 lt 001) enhanced the DPPH clearance and ∙OH clearance(Table 1) Considering the sound probiotic properties of theL plantarum HM2187491 [15] this strain was finally chosenfor the antibacterial evaluation

Interestingly no antimicrobial effect was observed usingAV extract and Aloe fermentation supernatant (fermentedby L plantarum HM2187491 without additional glucose)while the addition of 5 glucose conferred the Aloe fer-mentation supernatant a perfect inhibition to all the testedpathogens for example S typhimurium ATCC 13311 (inhibi-tion zone diameter 16mm) S enteritidis ATCC13076 (IZD

4 Mediators of Inflammation

Table 1 The antioxidant activity of the Aloe vera Aloe vera + B longum Aloe vera + L plantarum Aloe vera + L rhamnosus GG and Aloevera + S thermophilus

Group DPPH clearance () O2

∙minus clearance () ∙OH clearance () Fe2+ chelation () Reducing powers (mgL)Aloe vera 6346 plusmn 413 9234 plusmn 712 6823 plusmn 223 8351 plusmn 478 0244 plusmn 0032

Aloe vera + B longum 5673 plusmn 612 9428 plusmn 548 7429 plusmn 154lowastlowast 8234 plusmn 541 0246 plusmn 009

Aloe vera + L plantarum 8523 plusmn 423lowastlowast 9313 plusmn 423 7623 plusmn 672lowastlowast 8287 plusmn 623 0242 plusmn 013

Aloe vera + L rhamnosus 5523 plusmn 348 9212 plusmn 533 5987 plusmn 581lowastlowast 8723 plusmn 612 0239 plusmn 008

Aloe vera + S thermophilus 8213 plusmn 231lowastlowast 9319 plusmn 712 6152 plusmn 678lowastlowast 8323 plusmn 334 0251 plusmn 011

Note data are shown as the mean plusmn SD lowastlowast119875 lt 001

MRSAloe vera + glucose

Aloe vera

ND

ND

ND

ND

ND

ND

ND

ND

Inhi

bitio

n zo

ne (m

m)

10

20

30

40

P ac

nesA

TCC

1182

7

S a

ureu

sCO

Wan

1

S d

ysen

teria

e 301

L m

onoc

ytog

enes

ATCC

1911

1

E co

li4410

2

S fl

exne

ri AT

CC 1

2022

S en

terit

idis

ATCC

1307

6

S ty

phim

uriu

mAT

CC13

311

Figure 2 Antibacterial activities of L plantarum against selected foodborne pathogens cultured in MRS Aloe vera + glucose and Aloe veraData are shown as the mean plusmn SD

15mm) S flexneri ATCC 12022 (IZD 19mm) E coli 44102(IZD 11mm) L monocytogenes ATCC 19111 (IZD 20mm)S dysenteriae 301 (IZD 21mm) and S aureus COWAN1(IZD 20mm) (Figure 2) Most of all the Aloe fermentationsupernatant possessed the best antimicrobial effect onP acnesATCC 11827 (IZD 27mm) which leads to an accumulationof the skins natural oils bacteria and acne vulgaris [23]

33 The Anti-Inflammatory Activity of the Aloe FermentationSupernatant on Proinflammatory Cytokines During inflam-mation excess levels of cytokines (IL-1120573 IL-6 and TNF-120572)lead to the damaging of cells and tissues and the activationof macrophages in inflammation-associated diseases (egrheumatoid arthritis and chronic hepatitis) [24 25]

To check the functional components of AV and Aloe fer-mentation supernatant on the production of IL-1120573 IL-6 andTNF-120572 different concentrations of emodin (0 1 25 5 and10 120583gmL) polysaccharide (0 25 5 15 30 and 50120583gmL)AV extract (0 5 15 30 50 100 and 200120583gmL) andAloe fer-mentation supernatant (0 5 15 30 50 100 and 200 120583gmL)

were cocultured with RAW 2647 cells to test their toxicityon RAW 2647 cells and the results indicated that no obvioustoxicity was observed for all the tested materials and the lowconcentration of emodin (1120583gmL) showedpromoting effectson the growth of RAW 2647 cells (Figure 3)

Then low and high doses of the above drugs were deter-mined to evaluate the anti-inflammatory effects of Aloe fer-mentation supernatant according to the previous studies [120 22 26 27]The real-time PCR results indicated that all thetested emodin polysaccharide AV extract and Aloe fermen-tation supernatant had significantly reduced the inflamma-tory factor of IL-1120573 IL-6 and TNF-120572 in gene level except forthe polysaccharide at 15 120583gmL In protein level our ELISAresults showed that both the AV extract and Aloe fermenta-tion supernatant presented significantly inhibition on IL-1120573IL-6 and TNF-120572 production and it seemed that the inhibi-tion effect of AV extract was better than Aloe fermentationsupernatant However little inhibition effect of emodin andpolysaccharide on IL-1120573 and polysaccharide on IL-6 wasobserved (Figure 5)

Mediators of Inflammation 5

Emodin

lowastlowast

1 50 1025

Concentration (120583gmL)

05

06

07

08O

D570

(a)

Polysaccharide

30 501525 50

Concentration (120583gmL)

05

06

07

08

09

OD

570

(b)

Aloe veraAloe vera + L plantarum

05

06

07

08

09

OD

570

50 15 5030 200100

Concentration (120583gmL)

(c)

Figure 3 Different concentration of emodin (a) polysaccharide (b) Aloe vera and Aloe vera + L plantarum (c) on the cell viability of RAW2647 cells using MTT method Data are shown as the mean plusmn SD lowastlowast119875 lt 001

4 Discussion

Aswe all know the single target drug had been widely used invarious human diseases but often failed to cure the metabolicdiseases [28]Thus the whole-body systems biology was pro-posed and regarded the human and symbiotic microorgan-isms as a whole ldquosuperorganismrdquo [28 29] which led a moreand more reorganization by the scientific community to theChinese herb extracts and the human intestinal probiotics

Previously studies showed that both the AV [2 22] andprobiotics [25 30 31] had sound antioxidant and anti-inflammatory effects so we guessed that the combination ofthe AV and probiotics should possess better antioxidativeantibacterial and anti-inflammatory effects First we opti-mized the optimum growth temperature of the selected pro-biotics and found that the probiotics could grew well in AVextract without any additives (Figure 1) However no antimi-crobial effects of AV and Aloe fermentation were observedwhich was inconsistent with the previous studies [2 32]

(Figure 2) We guessed that the lack of glucose the substratefor probiotics to produce antimicrobial production of lacticacid and acetic acid was the cause of the antimicrobial failurefor Aloe fermentation therefore we added 5 glucose to AVextract and found that the added glucose not only signif-icantly increased the probiotics growth (Figure 1) but alsoconferred the Aloe fermentation supernatant sound antibac-terial effects on all the tested pathogens (Figure 2)

After a series of evaluations the L plantarumHM2187491was chosen and this strain conferred the Aloe fermentationsupernatant strong scavenging capacities of the DPPH (86)O2

∙minus (85) ∙OH (76) Fe2+ chelation (82) and reducingpowers (2425mgL) (Table 1) Moreover the Aloe fermen-tation fermented by L plantarum HM2187491 could inhibitall the growth of tested pathogens especially for the P acnesATCC 11827 (IZD 27mm)Therefore the sound inhibition ofthe growth ofP acnesATCC 11827 (causative bacteria for acnevulgaris) and antioxidant activitymade theAloe fermentationsupernatant a potential ingredient for cosmetic

6 Mediators of Inflammation

IL-1

120573 m

RNA

GA

PDH

mRN

A

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(a)

TNF-

a mRN

AG

APD

H m

RNA

0

10

20

30

40

lowastlowast

lowastlowast

lowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)

IL-6

mRN

AG

APD

H m

RNA

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 4 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the expression ofinflammatory genes (IL-1120573 TNF-120572 and IL-6) in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

As we know excess levels of IL-1120573 IL-6 and TNF-120572induced by activated inflammatory cells (eg eosinophilsmacrophages mononuclear phagocytes neutrophils) willlead to the damage of cells and tissues which eventually causethe inflammation-associated diseases for example rheuma-toid arthritis chronic hepatitis and macrophages play animportant role in regulating several immunopathologicalconditions and inducing overexpression of the proinflamma-tory mediators of the inflammatory process [24 25] So theRAW2647 cells were used in the present study to evaluate theanti-inflammatory effect of Aloe fermentation supernatantfermented by L plantarumHM2187491

The results indicated that both the emodin (anticanceractivity) [33] and the polysaccharide (immunity enhance-ment and reduction of oxidative injury) showed weakeranti-inflammatory effects compared to AV extract and Aloefermentation supernatant in mRNA level and protein level(Figures 4 and 5) Interestingly though theAloe fermentationsupernatant possessed a strong antioxidant effect (Figure 2)

its anti-inflammatory effect was inferior to the AV extractin both gene level and protein level Maybe some anti-inflammatory ingredients in AV extract were consumed byL plantarum HM2187491 and the metabolites of this strainmight contribute to the inflammatory response of RAW2647 cells Moreover in vivo work is needed because of theshortcomings of cell model for example it could not modelthe interaction between intestinal cells and microbiota Aspotential probiotics the L plantarum HM2187491 couldproduce bacteriocins offering advantages in colonization andcompetition in the gastrointestinal tract [34] and endowedthe AV extract a strong antioxidative effect and antimicrobialeffect on pathogens (especially for P acnes) In addition theL plantarumHM2187491 could remove the bitter taste of AVand our human results (119899 = 15) indicated that the Aloe fer-mentation supernatant had shortened the acne cure time andhad a perfect whitening effect (data unshown)

In conclusion the Aloe fermentation supernatantrsquos soundantimicrobial effect and anti-inflammatory effect together

Mediators of Inflammation 7

IL-1120573(p

gm

L)

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

lowastlowast

lowastlowast

lowastlowast lowastlowast

0

10

20

30

40

50

(a)

TNF-120572

(pg

mL)

0

500

1000

1500

lowastlowastlowastlowast lowastlowast

lowastlowast

lowastlowastlowastlowast lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

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+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)IL-6

(pg

mL)

lowastlowast

lowastlowast

lowastlowast lowastlowast lowastlowastlowastlowast

0

50

100

150

200

250

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 5 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the protein yields of IL-1120573TNF-120572 and IL-6 in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

with its intestinal health promoting effect when L plantarumHM2187491 enters host intestines indicate its potential useas functional foods or cosmetics to guard human intestinalhealth delaying senescence and preventing chronic diseases

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

This work was supported by grants from the National BasicResearch Program of China (2013CB531103) and NationalNatural Science Foundation of China (nos 8150336431560264 91339113 81270202 and 21461015) and the grantsfrom Jiangxi Province (nos 20151BAB205001 GJJ14218KJLD14010 and 20153BCB23035)

References

[1] Y-W Kim Y-J Jeong A-Y Kim et al ldquoLactobacillus brevisstrains from fermented Aloe vera survive gastroduodenal envi-ronment and suppress common food borne enteropathogensrdquoPLoS ONE vol 9 no 3 Article ID e90866 2014

[2] E V Christaki and P C Florou-Paneri ldquoAloe vera a plant formany usesrdquo Journal of Food Agriculture and Environment vol8 no 2 pp 245ndash249 2010

[3] S Rajasekaran K Ravi K Sivagnanam and S SubramanianldquoBeneficial effects of Aloe vera leaf gel extract on lipid profilestatus in rats with streptozotocin diabetesrdquo Clinical and Experi-mental Pharmacology and Physiology vol 33 no 3 pp 232ndash2372006

[4] S Shamim S W Ahmed and I Azhar ldquoAntifungal activity ofAlliumAloe and Solanum speciesrdquo Pharmaceutical Biology vol42 no 7 pp 491ndash498 2004

[5] M M Bottenberg G C Wall R L Harvey and S Habib ldquoOralaloe vera-induced hepatitisrdquo Annals of Pharmacotherapy vol41 no 10 pp 1740ndash1743 2007

[6] L Langmead RM Feakins S Goldthorpe et al ldquoRandomizeddouble-blind placebo-controlled trial of oral aloe vera gel foractive ulcerative colitisrdquoAlimentary Pharmacology ampTherapeu-tics vol 19 no 7 pp 739ndash747 2004

[7] S Li T Chen F Xu et al ldquoThe beneficial effect of exopolysac-charides from Bifidobacterium bifidum WBIN03 on microbialdiversity in mouse intestinerdquo Journal of the Science of Food andAgriculture vol 94 no 2 pp 256ndash264 2014

[8] T Chen S Li and H Wei ldquoAntibiotic resistance capability ofcultured human colonic microbiota growing in a chemostat

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

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Diabetes ResearchJournal of

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Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

2 Mediators of Inflammation

functional foods and potential oral vaccines [13] In ourprevious work [15] L plantarum HM2187491 was isolatedfrom human intestinal which could grow better at 37∘C in ananaerobic environment and could resist harsh conditions ofgastrointestinal tract (pH 25 and 030 bile salt) Moreoverthis strain could strongly adhere toHT-29 cells and reduce theadhesion of Shigella dysenteriae 2457 Staphylococcus aureusCowan1 Enterobacter sakazakii 45401 and Escherichia coli44102 to HT-29 cells and its long-term residence in miceintestine increased the number of lactobacilli and decreasedthe number of enterococci [15]

In the present study the tested probiotics (eg Bifi-dobacterium longumATCC15708 L plantarumHM218749 LrhamnosusGG and Streptococcus thermophilusG1) grew wellin AV substrate and conferred AV sound antioxidant andantibacterial effects and the L plantarum HM2187491 wasfinally chosen for its popularity in food fields and perfectbeneficial effects verified in our previous studies and thecell experiment was carried out to further verify the anti-inflammatory effects of the Aloe fermentation supernatant

2 Materials and Methods

21 Preparation of AV Extract and Aloe Fermentation Super-natant TheAV leaves were freshly harvested by cutting fromthe bottom and were washed thoroughly The edges wereremoved and the leaves were peeled to obtain the slimy andtransparent gel contents The gel was then mashed using apulper which was then centrifuged at 4000timesg for 10min toobtain the AV extract [1]

The probiotic Bifidobacterium longum ATCC15708 (iso-lated from the milk) L plantarum HM218749 (isolated fromhuman intestine) L rhamnosusGG (isolated from the milk)and Streptococcus thermophilus G1 (isolated from the milk)were cultured in de Man Rogosa Sharpe (MRS) medium (forLactobacilli and Bifidobacterium) and brainheart infusion(BHI) medium (for Streptococcus thermophilus G1) for 24 hand 108 cfumL of the strains was used as an inoculum to beadded to Aloe vera Aloe vera + 5 glucose or Aloe vera +5 glucose + 5 skimmed milk for preparing the Aloe fer-mentation supernatant which was incubated for 15 to 18 h at37∘C or 42∘C Finally the Aloe fermentation supernatant andAV were freeze-dried using vacuum freeze drier under withthe following conditions freeze drying at 02mbar vacuumminus40∘C for 48 h The lyophilized powder was stored at minus20∘Cfor further use

22 Cultivable Bacterial Counts Bacterial counts were enu-merated as previously reported [16] Diluted aliquots werereplica-plated onto BHI agar and MRS agar and then incu-bated anaerobically in the anaerobic system (full of 85 N

2

5H2 and 10CO

2) at 37∘Cor 42∘C for 24sim36 h followed by

counting colonies from plates showing 25ndash250 colonies [17]

23 Antioxidative and Antibacterial Activity of AV Extract andAloe Fermentation Supernatant For preparation of the Aloefermentation supernatant 10 of the probiotics were addedto AV extract and fermented for 15 to 18 h at 37∘C and thenthe Aloe fermentation cultures were centrifuged at 4000timesg

for 10min and the pellets in the bottom of the centrifugetubes (containing insoluble materials orand probiotics) weredeemed as the residual substance and the clear water insolu-ble substance was deemed as supernatant and the AV extractwas used as a control Then the clearance of the DPPHO2

∙minus ∙OH Fe2+ chelation and oxygen-reduction activitywere measured just as in [18] For antimicrobial activityovernight (12 h) cultures of the pathogenic microorganismsincluding Salmonella typhimurium ATCC 13311 S enteritidisATCC13076 S flexneri ATCC 12022 E coli 44102 Listeriamonocytogenes ATCC 19111 S dysenteriae 301 S aureusCowan1 and Propionibacterium acnes ATCC 11827 werespread on the surface of the LB agar plates and the culturesupernatant (200120583L) was loaded into an Oxford cup (outerdiameter 78 plusmn 01mm inner diameter 60 plusmn 01mm andheight 100plusmn01mm) which was placed on the surface of theagarThe size of the inhibition zone was measured until a for-mation of clear zone around the Oxford cupThe experimentwas carried out in duplicate [17]

24 Cell Viability Assay RAW 2647 cells (passages 8ndash20)were maintained in DMEM supplemented with 10 FBSand antibiotics (100UmL of penicillin and 100 120583gmL ofstreptomycin) at 37∘C in a 5 CO

2incubator Emodin

(Wuhan qi Boster Biological Technology co Ltd) polysac-charide (Wuhan qi Boster Biological Technology co Ltd)Aloe vera (lyophilized powder) and Aloe vera + L plantarum(lyophilized powder) were dissolved in phosphate-bufferedsaline (PBS) solution and their effect on cell viability of RAW2647 cells was determined by MTT assay [19]

RAW 2647 cells were plated at a density of 10000 perwell in 100 120583L of complete culture medium and treatedwith designed concentrations of emodin (0 1 25 5 and10 120583gmL) polysaccharide (0 25 5 15 30 and 50 120583gmL)Aloe vera (0 5 15 30 50 100 and 200120583gmL) and Aloe vera+ L plantarum (0 5 15 30 50 100 and 200120583gmL) in 96-wellmicrotiter plates for 36 h at 37∘C in a humidified incubatorControl cells treated with PBS served as the vehicle groupEach concentration of drugs was repeated in 5 wells Afterincubation for specified times MTT reagent (20120583L 5mgmL) was added to each well and incubated for 4 h After care-ful removal of theMTT solution 150 120583L of DMSOwas addedto each well The absorbance was recorded on a microplatereader at the wavelength of 570 nm

25 Anti-Inflammatory Evaluation of AV Extract and AloeFermentation Supernatant Using RT-PCR and ELISA RAW2647 cells were seeded into 24-well plates (5 times 104 cells perwell) for 24 h and were then washed with PBS Cells weretreatedwithwithout lipopolysaccharide (LPS 1 120583gmL each)emodin (25120583gmL and 10 120583gmL) polysaccharide (25120583gmL and 15 120583gmL) Aloe vera (5 120583gmL and 50 120583gmL) andAloe vera + L plantarum (5 120583gmL and 50120583gmL) for 24 hand then the cells and supernatants were harvested for Q-PCR and ELISA analysis respectively

For the evaluation of cytokine mRNA expression levelstotal RNAs from the RAW 2647 cells were prepared byadding TRIzol reagent (Gibco BRL Grand Island NY USA)according to the manufacturerrsquos protocol and 24 120583g of total

Mediators of Inflammation 3

B longumL plantarum

L rhamnosusS thermophilus

Lg C

FUm

L

30 37 4225

Temperature (∘C)

8

9

10

11

12

(a)

B longum L plantarum L rhamnosus S thermophilus

Lg C

FUm

L

Aloe veraAloe vera + glucoseAloe vera + glucose + skimmed milk

8

9

10

11

12

lowastlowast

lowastlowast lowastlowast lowastlowast lowastlowast

lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast

(b)

Figure 1 The suitable temperature (a) and fermentation substrates (b) for B longum L plantarum L rhamnosus GG and S thermophilusThe significant analysis was done between Aloe vera Aloe vera + glucose and Aloe vera + glucose + skimmed milk Data are shown as themean plusmn SD lowastlowast119875 lt 001

RNA from each group was reverse transcribed to cDNAusing a commercially available kit (Applied Biosystems)Quantitative real-time PCRwas performed with 7900HT fastreal-time PCR system (ABI Foster City CA) using 2 times SYBRGreen master mix (Bio-Rad) Forty cycles were conducted asfollows 95∘C for 30 s and 60∘C for 30 s preceded by 1minat 95∘C for polymerase activation with the following primers(Q-PCR IL-1120573 sense primer 51015840-GTGTCTTTCCCGTGG-ACCTTC1015840-31015840 antisense primer 51015840-TCATCTCGGAGCCTG-TAGTGC-31015840 Q-PCR TNF-120572 sense primer 51015840-GTGGAA-CTGGCAGAAGAGGCA antisense primer 51015840-AGAGGG-AGGCCATTTGGGAAC-31015840 Q-PCR IL-6 sense primer 51015840-GGAAATCGTGGAAATGAG-31015840 antisense primer 51015840-GCT-TAGGCATAACGCACT-31015840 Q-PCR GAPDH sense primer51015840-CTCGTGGAGTCTACTGGTGT-31015840 antisense primer 51015840-GTCATCATACTTGGCAGGTT-31015840)

The products of IL-1120573 TNF-120572 and IL-6 in cell super-natants were determined using the ELISA kit for IL-1120573(eBioscience) TNF-120572 (eBioscience) and IL-6 (eBioscience)

26 Data Analysis Data are reported as means plusmn SD andresults were analyzed using SPSS 130 software (SPSS IncChicago IL USA) by means of independent one-wayANOVA tests in each sampling point The differences amongthe three groups were assessed by means of the least signifi-cant difference (LSD) multiple comparison test (119875 lt 005 or001)

3 Results

31 Exploration of the Fermentation Conditions of Probi-otics To determine the optimum growth temperature of theselected probiotics the same initial inoculums of B longum(MRS medium) L plantarum (MRS medium) L rhamnosusLGG (MRS medium) and S thermophilus (BHI medium)

were inoculated in the corresponding media and cultured at25∘C 30∘C 37∘C and 42∘C and the results showed that theoptimum temperature for B longum L plantarum and Lrhamnosus LGG was 37∘C and S thermophilus received itsmaximum yield at 42∘C (Figure 1(a))

To further investigate the effects of different additives onthe growth of probiotics glucose and skimmed milk wereadded in the AV extract and the results indicated that all thetested strains grew well in AV extract with the initial pH 60(about 9 LgCFUmL Figure 1(b)) It seemed that the additionof 5 glucose and 5 glucose + 5 skimmed milk had sig-nificantly enhanced the biomass of B longum L plantarumand L rhamnosus GG (119886 lt 001) and the growth promotingeffect of the 5 glucose + 5 skimmed milk was muchbetter than 5 glucose (119887 lt 001) (Figure 1(b))

32 Antioxidative and Antibacterial Activity of Aloe Fermen-tation Supernatant To evaluate the additive of the probioticson the perfect antioxidant and antibacterial activities of AV[9 20ndash22] the AV + 5 glucose which gives us more choicesfor the future products was chosen for our next study

Compared with the AV the addition of probiotics madelittle change on O

2

∙minus clearance Fe2+ chelation and reductionactivity while the L plantarumHM2187491 had significantly(119875 lt 001) enhanced the DPPH clearance and ∙OH clearance(Table 1) Considering the sound probiotic properties of theL plantarum HM2187491 [15] this strain was finally chosenfor the antibacterial evaluation

Interestingly no antimicrobial effect was observed usingAV extract and Aloe fermentation supernatant (fermentedby L plantarum HM2187491 without additional glucose)while the addition of 5 glucose conferred the Aloe fer-mentation supernatant a perfect inhibition to all the testedpathogens for example S typhimurium ATCC 13311 (inhibi-tion zone diameter 16mm) S enteritidis ATCC13076 (IZD

4 Mediators of Inflammation

Table 1 The antioxidant activity of the Aloe vera Aloe vera + B longum Aloe vera + L plantarum Aloe vera + L rhamnosus GG and Aloevera + S thermophilus

Group DPPH clearance () O2

∙minus clearance () ∙OH clearance () Fe2+ chelation () Reducing powers (mgL)Aloe vera 6346 plusmn 413 9234 plusmn 712 6823 plusmn 223 8351 plusmn 478 0244 plusmn 0032

Aloe vera + B longum 5673 plusmn 612 9428 plusmn 548 7429 plusmn 154lowastlowast 8234 plusmn 541 0246 plusmn 009

Aloe vera + L plantarum 8523 plusmn 423lowastlowast 9313 plusmn 423 7623 plusmn 672lowastlowast 8287 plusmn 623 0242 plusmn 013

Aloe vera + L rhamnosus 5523 plusmn 348 9212 plusmn 533 5987 plusmn 581lowastlowast 8723 plusmn 612 0239 plusmn 008

Aloe vera + S thermophilus 8213 plusmn 231lowastlowast 9319 plusmn 712 6152 plusmn 678lowastlowast 8323 plusmn 334 0251 plusmn 011

Note data are shown as the mean plusmn SD lowastlowast119875 lt 001

MRSAloe vera + glucose

Aloe vera

ND

ND

ND

ND

ND

ND

ND

ND

Inhi

bitio

n zo

ne (m

m)

10

20

30

40

P ac

nesA

TCC

1182

7

S a

ureu

sCO

Wan

1

S d

ysen

teria

e 301

L m

onoc

ytog

enes

ATCC

1911

1

E co

li4410

2

S fl

exne

ri AT

CC 1

2022

S en

terit

idis

ATCC

1307

6

S ty

phim

uriu

mAT

CC13

311

Figure 2 Antibacterial activities of L plantarum against selected foodborne pathogens cultured in MRS Aloe vera + glucose and Aloe veraData are shown as the mean plusmn SD

15mm) S flexneri ATCC 12022 (IZD 19mm) E coli 44102(IZD 11mm) L monocytogenes ATCC 19111 (IZD 20mm)S dysenteriae 301 (IZD 21mm) and S aureus COWAN1(IZD 20mm) (Figure 2) Most of all the Aloe fermentationsupernatant possessed the best antimicrobial effect onP acnesATCC 11827 (IZD 27mm) which leads to an accumulationof the skins natural oils bacteria and acne vulgaris [23]

33 The Anti-Inflammatory Activity of the Aloe FermentationSupernatant on Proinflammatory Cytokines During inflam-mation excess levels of cytokines (IL-1120573 IL-6 and TNF-120572)lead to the damaging of cells and tissues and the activationof macrophages in inflammation-associated diseases (egrheumatoid arthritis and chronic hepatitis) [24 25]

To check the functional components of AV and Aloe fer-mentation supernatant on the production of IL-1120573 IL-6 andTNF-120572 different concentrations of emodin (0 1 25 5 and10 120583gmL) polysaccharide (0 25 5 15 30 and 50120583gmL)AV extract (0 5 15 30 50 100 and 200120583gmL) andAloe fer-mentation supernatant (0 5 15 30 50 100 and 200 120583gmL)

were cocultured with RAW 2647 cells to test their toxicityon RAW 2647 cells and the results indicated that no obvioustoxicity was observed for all the tested materials and the lowconcentration of emodin (1120583gmL) showedpromoting effectson the growth of RAW 2647 cells (Figure 3)

Then low and high doses of the above drugs were deter-mined to evaluate the anti-inflammatory effects of Aloe fer-mentation supernatant according to the previous studies [120 22 26 27]The real-time PCR results indicated that all thetested emodin polysaccharide AV extract and Aloe fermen-tation supernatant had significantly reduced the inflamma-tory factor of IL-1120573 IL-6 and TNF-120572 in gene level except forthe polysaccharide at 15 120583gmL In protein level our ELISAresults showed that both the AV extract and Aloe fermenta-tion supernatant presented significantly inhibition on IL-1120573IL-6 and TNF-120572 production and it seemed that the inhibi-tion effect of AV extract was better than Aloe fermentationsupernatant However little inhibition effect of emodin andpolysaccharide on IL-1120573 and polysaccharide on IL-6 wasobserved (Figure 5)

Mediators of Inflammation 5

Emodin

lowastlowast

1 50 1025

Concentration (120583gmL)

05

06

07

08O

D570

(a)

Polysaccharide

30 501525 50

Concentration (120583gmL)

05

06

07

08

09

OD

570

(b)

Aloe veraAloe vera + L plantarum

05

06

07

08

09

OD

570

50 15 5030 200100

Concentration (120583gmL)

(c)

Figure 3 Different concentration of emodin (a) polysaccharide (b) Aloe vera and Aloe vera + L plantarum (c) on the cell viability of RAW2647 cells using MTT method Data are shown as the mean plusmn SD lowastlowast119875 lt 001

4 Discussion

Aswe all know the single target drug had been widely used invarious human diseases but often failed to cure the metabolicdiseases [28]Thus the whole-body systems biology was pro-posed and regarded the human and symbiotic microorgan-isms as a whole ldquosuperorganismrdquo [28 29] which led a moreand more reorganization by the scientific community to theChinese herb extracts and the human intestinal probiotics

Previously studies showed that both the AV [2 22] andprobiotics [25 30 31] had sound antioxidant and anti-inflammatory effects so we guessed that the combination ofthe AV and probiotics should possess better antioxidativeantibacterial and anti-inflammatory effects First we opti-mized the optimum growth temperature of the selected pro-biotics and found that the probiotics could grew well in AVextract without any additives (Figure 1) However no antimi-crobial effects of AV and Aloe fermentation were observedwhich was inconsistent with the previous studies [2 32]

(Figure 2) We guessed that the lack of glucose the substratefor probiotics to produce antimicrobial production of lacticacid and acetic acid was the cause of the antimicrobial failurefor Aloe fermentation therefore we added 5 glucose to AVextract and found that the added glucose not only signif-icantly increased the probiotics growth (Figure 1) but alsoconferred the Aloe fermentation supernatant sound antibac-terial effects on all the tested pathogens (Figure 2)

After a series of evaluations the L plantarumHM2187491was chosen and this strain conferred the Aloe fermentationsupernatant strong scavenging capacities of the DPPH (86)O2

∙minus (85) ∙OH (76) Fe2+ chelation (82) and reducingpowers (2425mgL) (Table 1) Moreover the Aloe fermen-tation fermented by L plantarum HM2187491 could inhibitall the growth of tested pathogens especially for the P acnesATCC 11827 (IZD 27mm)Therefore the sound inhibition ofthe growth ofP acnesATCC 11827 (causative bacteria for acnevulgaris) and antioxidant activitymade theAloe fermentationsupernatant a potential ingredient for cosmetic

6 Mediators of Inflammation

IL-1

120573 m

RNA

GA

PDH

mRN

A

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(a)

TNF-

a mRN

AG

APD

H m

RNA

0

10

20

30

40

lowastlowast

lowastlowast

lowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)

IL-6

mRN

AG

APD

H m

RNA

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 4 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the expression ofinflammatory genes (IL-1120573 TNF-120572 and IL-6) in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

As we know excess levels of IL-1120573 IL-6 and TNF-120572induced by activated inflammatory cells (eg eosinophilsmacrophages mononuclear phagocytes neutrophils) willlead to the damage of cells and tissues which eventually causethe inflammation-associated diseases for example rheuma-toid arthritis chronic hepatitis and macrophages play animportant role in regulating several immunopathologicalconditions and inducing overexpression of the proinflamma-tory mediators of the inflammatory process [24 25] So theRAW2647 cells were used in the present study to evaluate theanti-inflammatory effect of Aloe fermentation supernatantfermented by L plantarumHM2187491

The results indicated that both the emodin (anticanceractivity) [33] and the polysaccharide (immunity enhance-ment and reduction of oxidative injury) showed weakeranti-inflammatory effects compared to AV extract and Aloefermentation supernatant in mRNA level and protein level(Figures 4 and 5) Interestingly though theAloe fermentationsupernatant possessed a strong antioxidant effect (Figure 2)

its anti-inflammatory effect was inferior to the AV extractin both gene level and protein level Maybe some anti-inflammatory ingredients in AV extract were consumed byL plantarum HM2187491 and the metabolites of this strainmight contribute to the inflammatory response of RAW2647 cells Moreover in vivo work is needed because of theshortcomings of cell model for example it could not modelthe interaction between intestinal cells and microbiota Aspotential probiotics the L plantarum HM2187491 couldproduce bacteriocins offering advantages in colonization andcompetition in the gastrointestinal tract [34] and endowedthe AV extract a strong antioxidative effect and antimicrobialeffect on pathogens (especially for P acnes) In addition theL plantarumHM2187491 could remove the bitter taste of AVand our human results (119899 = 15) indicated that the Aloe fer-mentation supernatant had shortened the acne cure time andhad a perfect whitening effect (data unshown)

In conclusion the Aloe fermentation supernatantrsquos soundantimicrobial effect and anti-inflammatory effect together

Mediators of Inflammation 7

IL-1120573(p

gm

L)

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

lowastlowast

lowastlowast

lowastlowast lowastlowast

0

10

20

30

40

50

(a)

TNF-120572

(pg

mL)

0

500

1000

1500

lowastlowastlowastlowast lowastlowast

lowastlowast

lowastlowastlowastlowast lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)IL-6

(pg

mL)

lowastlowast

lowastlowast

lowastlowast lowastlowast lowastlowastlowastlowast

0

50

100

150

200

250

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 5 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the protein yields of IL-1120573TNF-120572 and IL-6 in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

with its intestinal health promoting effect when L plantarumHM2187491 enters host intestines indicate its potential useas functional foods or cosmetics to guard human intestinalhealth delaying senescence and preventing chronic diseases

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

This work was supported by grants from the National BasicResearch Program of China (2013CB531103) and NationalNatural Science Foundation of China (nos 8150336431560264 91339113 81270202 and 21461015) and the grantsfrom Jiangxi Province (nos 20151BAB205001 GJJ14218KJLD14010 and 20153BCB23035)

References

[1] Y-W Kim Y-J Jeong A-Y Kim et al ldquoLactobacillus brevisstrains from fermented Aloe vera survive gastroduodenal envi-ronment and suppress common food borne enteropathogensrdquoPLoS ONE vol 9 no 3 Article ID e90866 2014

[2] E V Christaki and P C Florou-Paneri ldquoAloe vera a plant formany usesrdquo Journal of Food Agriculture and Environment vol8 no 2 pp 245ndash249 2010

[3] S Rajasekaran K Ravi K Sivagnanam and S SubramanianldquoBeneficial effects of Aloe vera leaf gel extract on lipid profilestatus in rats with streptozotocin diabetesrdquo Clinical and Experi-mental Pharmacology and Physiology vol 33 no 3 pp 232ndash2372006

[4] S Shamim S W Ahmed and I Azhar ldquoAntifungal activity ofAlliumAloe and Solanum speciesrdquo Pharmaceutical Biology vol42 no 7 pp 491ndash498 2004

[5] M M Bottenberg G C Wall R L Harvey and S Habib ldquoOralaloe vera-induced hepatitisrdquo Annals of Pharmacotherapy vol41 no 10 pp 1740ndash1743 2007

[6] L Langmead RM Feakins S Goldthorpe et al ldquoRandomizeddouble-blind placebo-controlled trial of oral aloe vera gel foractive ulcerative colitisrdquoAlimentary Pharmacology ampTherapeu-tics vol 19 no 7 pp 739ndash747 2004

[7] S Li T Chen F Xu et al ldquoThe beneficial effect of exopolysac-charides from Bifidobacterium bifidum WBIN03 on microbialdiversity in mouse intestinerdquo Journal of the Science of Food andAgriculture vol 94 no 2 pp 256ndash264 2014

[8] T Chen S Li and H Wei ldquoAntibiotic resistance capability ofcultured human colonic microbiota growing in a chemostat

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

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Diabetes ResearchJournal of

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Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Mediators of Inflammation 3

B longumL plantarum

L rhamnosusS thermophilus

Lg C

FUm

L

30 37 4225

Temperature (∘C)

8

9

10

11

12

(a)

B longum L plantarum L rhamnosus S thermophilus

Lg C

FUm

L

Aloe veraAloe vera + glucoseAloe vera + glucose + skimmed milk

8

9

10

11

12

lowastlowast

lowastlowast lowastlowast lowastlowast lowastlowast

lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast lowastlowast

(b)

Figure 1 The suitable temperature (a) and fermentation substrates (b) for B longum L plantarum L rhamnosus GG and S thermophilusThe significant analysis was done between Aloe vera Aloe vera + glucose and Aloe vera + glucose + skimmed milk Data are shown as themean plusmn SD lowastlowast119875 lt 001

RNA from each group was reverse transcribed to cDNAusing a commercially available kit (Applied Biosystems)Quantitative real-time PCRwas performed with 7900HT fastreal-time PCR system (ABI Foster City CA) using 2 times SYBRGreen master mix (Bio-Rad) Forty cycles were conducted asfollows 95∘C for 30 s and 60∘C for 30 s preceded by 1minat 95∘C for polymerase activation with the following primers(Q-PCR IL-1120573 sense primer 51015840-GTGTCTTTCCCGTGG-ACCTTC1015840-31015840 antisense primer 51015840-TCATCTCGGAGCCTG-TAGTGC-31015840 Q-PCR TNF-120572 sense primer 51015840-GTGGAA-CTGGCAGAAGAGGCA antisense primer 51015840-AGAGGG-AGGCCATTTGGGAAC-31015840 Q-PCR IL-6 sense primer 51015840-GGAAATCGTGGAAATGAG-31015840 antisense primer 51015840-GCT-TAGGCATAACGCACT-31015840 Q-PCR GAPDH sense primer51015840-CTCGTGGAGTCTACTGGTGT-31015840 antisense primer 51015840-GTCATCATACTTGGCAGGTT-31015840)

The products of IL-1120573 TNF-120572 and IL-6 in cell super-natants were determined using the ELISA kit for IL-1120573(eBioscience) TNF-120572 (eBioscience) and IL-6 (eBioscience)

26 Data Analysis Data are reported as means plusmn SD andresults were analyzed using SPSS 130 software (SPSS IncChicago IL USA) by means of independent one-wayANOVA tests in each sampling point The differences amongthe three groups were assessed by means of the least signifi-cant difference (LSD) multiple comparison test (119875 lt 005 or001)

3 Results

31 Exploration of the Fermentation Conditions of Probi-otics To determine the optimum growth temperature of theselected probiotics the same initial inoculums of B longum(MRS medium) L plantarum (MRS medium) L rhamnosusLGG (MRS medium) and S thermophilus (BHI medium)

were inoculated in the corresponding media and cultured at25∘C 30∘C 37∘C and 42∘C and the results showed that theoptimum temperature for B longum L plantarum and Lrhamnosus LGG was 37∘C and S thermophilus received itsmaximum yield at 42∘C (Figure 1(a))

To further investigate the effects of different additives onthe growth of probiotics glucose and skimmed milk wereadded in the AV extract and the results indicated that all thetested strains grew well in AV extract with the initial pH 60(about 9 LgCFUmL Figure 1(b)) It seemed that the additionof 5 glucose and 5 glucose + 5 skimmed milk had sig-nificantly enhanced the biomass of B longum L plantarumand L rhamnosus GG (119886 lt 001) and the growth promotingeffect of the 5 glucose + 5 skimmed milk was muchbetter than 5 glucose (119887 lt 001) (Figure 1(b))

32 Antioxidative and Antibacterial Activity of Aloe Fermen-tation Supernatant To evaluate the additive of the probioticson the perfect antioxidant and antibacterial activities of AV[9 20ndash22] the AV + 5 glucose which gives us more choicesfor the future products was chosen for our next study

Compared with the AV the addition of probiotics madelittle change on O

2

∙minus clearance Fe2+ chelation and reductionactivity while the L plantarumHM2187491 had significantly(119875 lt 001) enhanced the DPPH clearance and ∙OH clearance(Table 1) Considering the sound probiotic properties of theL plantarum HM2187491 [15] this strain was finally chosenfor the antibacterial evaluation

Interestingly no antimicrobial effect was observed usingAV extract and Aloe fermentation supernatant (fermentedby L plantarum HM2187491 without additional glucose)while the addition of 5 glucose conferred the Aloe fer-mentation supernatant a perfect inhibition to all the testedpathogens for example S typhimurium ATCC 13311 (inhibi-tion zone diameter 16mm) S enteritidis ATCC13076 (IZD

4 Mediators of Inflammation

Table 1 The antioxidant activity of the Aloe vera Aloe vera + B longum Aloe vera + L plantarum Aloe vera + L rhamnosus GG and Aloevera + S thermophilus

Group DPPH clearance () O2

∙minus clearance () ∙OH clearance () Fe2+ chelation () Reducing powers (mgL)Aloe vera 6346 plusmn 413 9234 plusmn 712 6823 plusmn 223 8351 plusmn 478 0244 plusmn 0032

Aloe vera + B longum 5673 plusmn 612 9428 plusmn 548 7429 plusmn 154lowastlowast 8234 plusmn 541 0246 plusmn 009

Aloe vera + L plantarum 8523 plusmn 423lowastlowast 9313 plusmn 423 7623 plusmn 672lowastlowast 8287 plusmn 623 0242 plusmn 013

Aloe vera + L rhamnosus 5523 plusmn 348 9212 plusmn 533 5987 plusmn 581lowastlowast 8723 plusmn 612 0239 plusmn 008

Aloe vera + S thermophilus 8213 plusmn 231lowastlowast 9319 plusmn 712 6152 plusmn 678lowastlowast 8323 plusmn 334 0251 plusmn 011

Note data are shown as the mean plusmn SD lowastlowast119875 lt 001

MRSAloe vera + glucose

Aloe vera

ND

ND

ND

ND

ND

ND

ND

ND

Inhi

bitio

n zo

ne (m

m)

10

20

30

40

P ac

nesA

TCC

1182

7

S a

ureu

sCO

Wan

1

S d

ysen

teria

e 301

L m

onoc

ytog

enes

ATCC

1911

1

E co

li4410

2

S fl

exne

ri AT

CC 1

2022

S en

terit

idis

ATCC

1307

6

S ty

phim

uriu

mAT

CC13

311

Figure 2 Antibacterial activities of L plantarum against selected foodborne pathogens cultured in MRS Aloe vera + glucose and Aloe veraData are shown as the mean plusmn SD

15mm) S flexneri ATCC 12022 (IZD 19mm) E coli 44102(IZD 11mm) L monocytogenes ATCC 19111 (IZD 20mm)S dysenteriae 301 (IZD 21mm) and S aureus COWAN1(IZD 20mm) (Figure 2) Most of all the Aloe fermentationsupernatant possessed the best antimicrobial effect onP acnesATCC 11827 (IZD 27mm) which leads to an accumulationof the skins natural oils bacteria and acne vulgaris [23]

33 The Anti-Inflammatory Activity of the Aloe FermentationSupernatant on Proinflammatory Cytokines During inflam-mation excess levels of cytokines (IL-1120573 IL-6 and TNF-120572)lead to the damaging of cells and tissues and the activationof macrophages in inflammation-associated diseases (egrheumatoid arthritis and chronic hepatitis) [24 25]

To check the functional components of AV and Aloe fer-mentation supernatant on the production of IL-1120573 IL-6 andTNF-120572 different concentrations of emodin (0 1 25 5 and10 120583gmL) polysaccharide (0 25 5 15 30 and 50120583gmL)AV extract (0 5 15 30 50 100 and 200120583gmL) andAloe fer-mentation supernatant (0 5 15 30 50 100 and 200 120583gmL)

were cocultured with RAW 2647 cells to test their toxicityon RAW 2647 cells and the results indicated that no obvioustoxicity was observed for all the tested materials and the lowconcentration of emodin (1120583gmL) showedpromoting effectson the growth of RAW 2647 cells (Figure 3)

Then low and high doses of the above drugs were deter-mined to evaluate the anti-inflammatory effects of Aloe fer-mentation supernatant according to the previous studies [120 22 26 27]The real-time PCR results indicated that all thetested emodin polysaccharide AV extract and Aloe fermen-tation supernatant had significantly reduced the inflamma-tory factor of IL-1120573 IL-6 and TNF-120572 in gene level except forthe polysaccharide at 15 120583gmL In protein level our ELISAresults showed that both the AV extract and Aloe fermenta-tion supernatant presented significantly inhibition on IL-1120573IL-6 and TNF-120572 production and it seemed that the inhibi-tion effect of AV extract was better than Aloe fermentationsupernatant However little inhibition effect of emodin andpolysaccharide on IL-1120573 and polysaccharide on IL-6 wasobserved (Figure 5)

Mediators of Inflammation 5

Emodin

lowastlowast

1 50 1025

Concentration (120583gmL)

05

06

07

08O

D570

(a)

Polysaccharide

30 501525 50

Concentration (120583gmL)

05

06

07

08

09

OD

570

(b)

Aloe veraAloe vera + L plantarum

05

06

07

08

09

OD

570

50 15 5030 200100

Concentration (120583gmL)

(c)

Figure 3 Different concentration of emodin (a) polysaccharide (b) Aloe vera and Aloe vera + L plantarum (c) on the cell viability of RAW2647 cells using MTT method Data are shown as the mean plusmn SD lowastlowast119875 lt 001

4 Discussion

Aswe all know the single target drug had been widely used invarious human diseases but often failed to cure the metabolicdiseases [28]Thus the whole-body systems biology was pro-posed and regarded the human and symbiotic microorgan-isms as a whole ldquosuperorganismrdquo [28 29] which led a moreand more reorganization by the scientific community to theChinese herb extracts and the human intestinal probiotics

Previously studies showed that both the AV [2 22] andprobiotics [25 30 31] had sound antioxidant and anti-inflammatory effects so we guessed that the combination ofthe AV and probiotics should possess better antioxidativeantibacterial and anti-inflammatory effects First we opti-mized the optimum growth temperature of the selected pro-biotics and found that the probiotics could grew well in AVextract without any additives (Figure 1) However no antimi-crobial effects of AV and Aloe fermentation were observedwhich was inconsistent with the previous studies [2 32]

(Figure 2) We guessed that the lack of glucose the substratefor probiotics to produce antimicrobial production of lacticacid and acetic acid was the cause of the antimicrobial failurefor Aloe fermentation therefore we added 5 glucose to AVextract and found that the added glucose not only signif-icantly increased the probiotics growth (Figure 1) but alsoconferred the Aloe fermentation supernatant sound antibac-terial effects on all the tested pathogens (Figure 2)

After a series of evaluations the L plantarumHM2187491was chosen and this strain conferred the Aloe fermentationsupernatant strong scavenging capacities of the DPPH (86)O2

∙minus (85) ∙OH (76) Fe2+ chelation (82) and reducingpowers (2425mgL) (Table 1) Moreover the Aloe fermen-tation fermented by L plantarum HM2187491 could inhibitall the growth of tested pathogens especially for the P acnesATCC 11827 (IZD 27mm)Therefore the sound inhibition ofthe growth ofP acnesATCC 11827 (causative bacteria for acnevulgaris) and antioxidant activitymade theAloe fermentationsupernatant a potential ingredient for cosmetic

6 Mediators of Inflammation

IL-1

120573 m

RNA

GA

PDH

mRN

A

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(a)

TNF-

a mRN

AG

APD

H m

RNA

0

10

20

30

40

lowastlowast

lowastlowast

lowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)

IL-6

mRN

AG

APD

H m

RNA

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 4 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the expression ofinflammatory genes (IL-1120573 TNF-120572 and IL-6) in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

As we know excess levels of IL-1120573 IL-6 and TNF-120572induced by activated inflammatory cells (eg eosinophilsmacrophages mononuclear phagocytes neutrophils) willlead to the damage of cells and tissues which eventually causethe inflammation-associated diseases for example rheuma-toid arthritis chronic hepatitis and macrophages play animportant role in regulating several immunopathologicalconditions and inducing overexpression of the proinflamma-tory mediators of the inflammatory process [24 25] So theRAW2647 cells were used in the present study to evaluate theanti-inflammatory effect of Aloe fermentation supernatantfermented by L plantarumHM2187491

The results indicated that both the emodin (anticanceractivity) [33] and the polysaccharide (immunity enhance-ment and reduction of oxidative injury) showed weakeranti-inflammatory effects compared to AV extract and Aloefermentation supernatant in mRNA level and protein level(Figures 4 and 5) Interestingly though theAloe fermentationsupernatant possessed a strong antioxidant effect (Figure 2)

its anti-inflammatory effect was inferior to the AV extractin both gene level and protein level Maybe some anti-inflammatory ingredients in AV extract were consumed byL plantarum HM2187491 and the metabolites of this strainmight contribute to the inflammatory response of RAW2647 cells Moreover in vivo work is needed because of theshortcomings of cell model for example it could not modelthe interaction between intestinal cells and microbiota Aspotential probiotics the L plantarum HM2187491 couldproduce bacteriocins offering advantages in colonization andcompetition in the gastrointestinal tract [34] and endowedthe AV extract a strong antioxidative effect and antimicrobialeffect on pathogens (especially for P acnes) In addition theL plantarumHM2187491 could remove the bitter taste of AVand our human results (119899 = 15) indicated that the Aloe fer-mentation supernatant had shortened the acne cure time andhad a perfect whitening effect (data unshown)

In conclusion the Aloe fermentation supernatantrsquos soundantimicrobial effect and anti-inflammatory effect together

Mediators of Inflammation 7

IL-1120573(p

gm

L)

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

lowastlowast

lowastlowast

lowastlowast lowastlowast

0

10

20

30

40

50

(a)

TNF-120572

(pg

mL)

0

500

1000

1500

lowastlowastlowastlowast lowastlowast

lowastlowast

lowastlowastlowastlowast lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)IL-6

(pg

mL)

lowastlowast

lowastlowast

lowastlowast lowastlowast lowastlowastlowastlowast

0

50

100

150

200

250

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 5 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the protein yields of IL-1120573TNF-120572 and IL-6 in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

with its intestinal health promoting effect when L plantarumHM2187491 enters host intestines indicate its potential useas functional foods or cosmetics to guard human intestinalhealth delaying senescence and preventing chronic diseases

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

This work was supported by grants from the National BasicResearch Program of China (2013CB531103) and NationalNatural Science Foundation of China (nos 8150336431560264 91339113 81270202 and 21461015) and the grantsfrom Jiangxi Province (nos 20151BAB205001 GJJ14218KJLD14010 and 20153BCB23035)

References

[1] Y-W Kim Y-J Jeong A-Y Kim et al ldquoLactobacillus brevisstrains from fermented Aloe vera survive gastroduodenal envi-ronment and suppress common food borne enteropathogensrdquoPLoS ONE vol 9 no 3 Article ID e90866 2014

[2] E V Christaki and P C Florou-Paneri ldquoAloe vera a plant formany usesrdquo Journal of Food Agriculture and Environment vol8 no 2 pp 245ndash249 2010

[3] S Rajasekaran K Ravi K Sivagnanam and S SubramanianldquoBeneficial effects of Aloe vera leaf gel extract on lipid profilestatus in rats with streptozotocin diabetesrdquo Clinical and Experi-mental Pharmacology and Physiology vol 33 no 3 pp 232ndash2372006

[4] S Shamim S W Ahmed and I Azhar ldquoAntifungal activity ofAlliumAloe and Solanum speciesrdquo Pharmaceutical Biology vol42 no 7 pp 491ndash498 2004

[5] M M Bottenberg G C Wall R L Harvey and S Habib ldquoOralaloe vera-induced hepatitisrdquo Annals of Pharmacotherapy vol41 no 10 pp 1740ndash1743 2007

[6] L Langmead RM Feakins S Goldthorpe et al ldquoRandomizeddouble-blind placebo-controlled trial of oral aloe vera gel foractive ulcerative colitisrdquoAlimentary Pharmacology ampTherapeu-tics vol 19 no 7 pp 739ndash747 2004

[7] S Li T Chen F Xu et al ldquoThe beneficial effect of exopolysac-charides from Bifidobacterium bifidum WBIN03 on microbialdiversity in mouse intestinerdquo Journal of the Science of Food andAgriculture vol 94 no 2 pp 256ndash264 2014

[8] T Chen S Li and H Wei ldquoAntibiotic resistance capability ofcultured human colonic microbiota growing in a chemostat

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

4 Mediators of Inflammation

Table 1 The antioxidant activity of the Aloe vera Aloe vera + B longum Aloe vera + L plantarum Aloe vera + L rhamnosus GG and Aloevera + S thermophilus

Group DPPH clearance () O2

∙minus clearance () ∙OH clearance () Fe2+ chelation () Reducing powers (mgL)Aloe vera 6346 plusmn 413 9234 plusmn 712 6823 plusmn 223 8351 plusmn 478 0244 plusmn 0032

Aloe vera + B longum 5673 plusmn 612 9428 plusmn 548 7429 plusmn 154lowastlowast 8234 plusmn 541 0246 plusmn 009

Aloe vera + L plantarum 8523 plusmn 423lowastlowast 9313 plusmn 423 7623 plusmn 672lowastlowast 8287 plusmn 623 0242 plusmn 013

Aloe vera + L rhamnosus 5523 plusmn 348 9212 plusmn 533 5987 plusmn 581lowastlowast 8723 plusmn 612 0239 plusmn 008

Aloe vera + S thermophilus 8213 plusmn 231lowastlowast 9319 plusmn 712 6152 plusmn 678lowastlowast 8323 plusmn 334 0251 plusmn 011

Note data are shown as the mean plusmn SD lowastlowast119875 lt 001

MRSAloe vera + glucose

Aloe vera

ND

ND

ND

ND

ND

ND

ND

ND

Inhi

bitio

n zo

ne (m

m)

10

20

30

40

P ac

nesA

TCC

1182

7

S a

ureu

sCO

Wan

1

S d

ysen

teria

e 301

L m

onoc

ytog

enes

ATCC

1911

1

E co

li4410

2

S fl

exne

ri AT

CC 1

2022

S en

terit

idis

ATCC

1307

6

S ty

phim

uriu

mAT

CC13

311

Figure 2 Antibacterial activities of L plantarum against selected foodborne pathogens cultured in MRS Aloe vera + glucose and Aloe veraData are shown as the mean plusmn SD

15mm) S flexneri ATCC 12022 (IZD 19mm) E coli 44102(IZD 11mm) L monocytogenes ATCC 19111 (IZD 20mm)S dysenteriae 301 (IZD 21mm) and S aureus COWAN1(IZD 20mm) (Figure 2) Most of all the Aloe fermentationsupernatant possessed the best antimicrobial effect onP acnesATCC 11827 (IZD 27mm) which leads to an accumulationof the skins natural oils bacteria and acne vulgaris [23]

33 The Anti-Inflammatory Activity of the Aloe FermentationSupernatant on Proinflammatory Cytokines During inflam-mation excess levels of cytokines (IL-1120573 IL-6 and TNF-120572)lead to the damaging of cells and tissues and the activationof macrophages in inflammation-associated diseases (egrheumatoid arthritis and chronic hepatitis) [24 25]

To check the functional components of AV and Aloe fer-mentation supernatant on the production of IL-1120573 IL-6 andTNF-120572 different concentrations of emodin (0 1 25 5 and10 120583gmL) polysaccharide (0 25 5 15 30 and 50120583gmL)AV extract (0 5 15 30 50 100 and 200120583gmL) andAloe fer-mentation supernatant (0 5 15 30 50 100 and 200 120583gmL)

were cocultured with RAW 2647 cells to test their toxicityon RAW 2647 cells and the results indicated that no obvioustoxicity was observed for all the tested materials and the lowconcentration of emodin (1120583gmL) showedpromoting effectson the growth of RAW 2647 cells (Figure 3)

Then low and high doses of the above drugs were deter-mined to evaluate the anti-inflammatory effects of Aloe fer-mentation supernatant according to the previous studies [120 22 26 27]The real-time PCR results indicated that all thetested emodin polysaccharide AV extract and Aloe fermen-tation supernatant had significantly reduced the inflamma-tory factor of IL-1120573 IL-6 and TNF-120572 in gene level except forthe polysaccharide at 15 120583gmL In protein level our ELISAresults showed that both the AV extract and Aloe fermenta-tion supernatant presented significantly inhibition on IL-1120573IL-6 and TNF-120572 production and it seemed that the inhibi-tion effect of AV extract was better than Aloe fermentationsupernatant However little inhibition effect of emodin andpolysaccharide on IL-1120573 and polysaccharide on IL-6 wasobserved (Figure 5)

Mediators of Inflammation 5

Emodin

lowastlowast

1 50 1025

Concentration (120583gmL)

05

06

07

08O

D570

(a)

Polysaccharide

30 501525 50

Concentration (120583gmL)

05

06

07

08

09

OD

570

(b)

Aloe veraAloe vera + L plantarum

05

06

07

08

09

OD

570

50 15 5030 200100

Concentration (120583gmL)

(c)

Figure 3 Different concentration of emodin (a) polysaccharide (b) Aloe vera and Aloe vera + L plantarum (c) on the cell viability of RAW2647 cells using MTT method Data are shown as the mean plusmn SD lowastlowast119875 lt 001

4 Discussion

Aswe all know the single target drug had been widely used invarious human diseases but often failed to cure the metabolicdiseases [28]Thus the whole-body systems biology was pro-posed and regarded the human and symbiotic microorgan-isms as a whole ldquosuperorganismrdquo [28 29] which led a moreand more reorganization by the scientific community to theChinese herb extracts and the human intestinal probiotics

Previously studies showed that both the AV [2 22] andprobiotics [25 30 31] had sound antioxidant and anti-inflammatory effects so we guessed that the combination ofthe AV and probiotics should possess better antioxidativeantibacterial and anti-inflammatory effects First we opti-mized the optimum growth temperature of the selected pro-biotics and found that the probiotics could grew well in AVextract without any additives (Figure 1) However no antimi-crobial effects of AV and Aloe fermentation were observedwhich was inconsistent with the previous studies [2 32]

(Figure 2) We guessed that the lack of glucose the substratefor probiotics to produce antimicrobial production of lacticacid and acetic acid was the cause of the antimicrobial failurefor Aloe fermentation therefore we added 5 glucose to AVextract and found that the added glucose not only signif-icantly increased the probiotics growth (Figure 1) but alsoconferred the Aloe fermentation supernatant sound antibac-terial effects on all the tested pathogens (Figure 2)

After a series of evaluations the L plantarumHM2187491was chosen and this strain conferred the Aloe fermentationsupernatant strong scavenging capacities of the DPPH (86)O2

∙minus (85) ∙OH (76) Fe2+ chelation (82) and reducingpowers (2425mgL) (Table 1) Moreover the Aloe fermen-tation fermented by L plantarum HM2187491 could inhibitall the growth of tested pathogens especially for the P acnesATCC 11827 (IZD 27mm)Therefore the sound inhibition ofthe growth ofP acnesATCC 11827 (causative bacteria for acnevulgaris) and antioxidant activitymade theAloe fermentationsupernatant a potential ingredient for cosmetic

6 Mediators of Inflammation

IL-1

120573 m

RNA

GA

PDH

mRN

A

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(a)

TNF-

a mRN

AG

APD

H m

RNA

0

10

20

30

40

lowastlowast

lowastlowast

lowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)

IL-6

mRN

AG

APD

H m

RNA

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 4 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the expression ofinflammatory genes (IL-1120573 TNF-120572 and IL-6) in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

As we know excess levels of IL-1120573 IL-6 and TNF-120572induced by activated inflammatory cells (eg eosinophilsmacrophages mononuclear phagocytes neutrophils) willlead to the damage of cells and tissues which eventually causethe inflammation-associated diseases for example rheuma-toid arthritis chronic hepatitis and macrophages play animportant role in regulating several immunopathologicalconditions and inducing overexpression of the proinflamma-tory mediators of the inflammatory process [24 25] So theRAW2647 cells were used in the present study to evaluate theanti-inflammatory effect of Aloe fermentation supernatantfermented by L plantarumHM2187491

The results indicated that both the emodin (anticanceractivity) [33] and the polysaccharide (immunity enhance-ment and reduction of oxidative injury) showed weakeranti-inflammatory effects compared to AV extract and Aloefermentation supernatant in mRNA level and protein level(Figures 4 and 5) Interestingly though theAloe fermentationsupernatant possessed a strong antioxidant effect (Figure 2)

its anti-inflammatory effect was inferior to the AV extractin both gene level and protein level Maybe some anti-inflammatory ingredients in AV extract were consumed byL plantarum HM2187491 and the metabolites of this strainmight contribute to the inflammatory response of RAW2647 cells Moreover in vivo work is needed because of theshortcomings of cell model for example it could not modelthe interaction between intestinal cells and microbiota Aspotential probiotics the L plantarum HM2187491 couldproduce bacteriocins offering advantages in colonization andcompetition in the gastrointestinal tract [34] and endowedthe AV extract a strong antioxidative effect and antimicrobialeffect on pathogens (especially for P acnes) In addition theL plantarumHM2187491 could remove the bitter taste of AVand our human results (119899 = 15) indicated that the Aloe fer-mentation supernatant had shortened the acne cure time andhad a perfect whitening effect (data unshown)

In conclusion the Aloe fermentation supernatantrsquos soundantimicrobial effect and anti-inflammatory effect together

Mediators of Inflammation 7

IL-1120573(p

gm

L)

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

lowastlowast

lowastlowast

lowastlowast lowastlowast

0

10

20

30

40

50

(a)

TNF-120572

(pg

mL)

0

500

1000

1500

lowastlowastlowastlowast lowastlowast

lowastlowast

lowastlowastlowastlowast lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)IL-6

(pg

mL)

lowastlowast

lowastlowast

lowastlowast lowastlowast lowastlowastlowastlowast

0

50

100

150

200

250

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 5 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the protein yields of IL-1120573TNF-120572 and IL-6 in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

with its intestinal health promoting effect when L plantarumHM2187491 enters host intestines indicate its potential useas functional foods or cosmetics to guard human intestinalhealth delaying senescence and preventing chronic diseases

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

This work was supported by grants from the National BasicResearch Program of China (2013CB531103) and NationalNatural Science Foundation of China (nos 8150336431560264 91339113 81270202 and 21461015) and the grantsfrom Jiangxi Province (nos 20151BAB205001 GJJ14218KJLD14010 and 20153BCB23035)

References

[1] Y-W Kim Y-J Jeong A-Y Kim et al ldquoLactobacillus brevisstrains from fermented Aloe vera survive gastroduodenal envi-ronment and suppress common food borne enteropathogensrdquoPLoS ONE vol 9 no 3 Article ID e90866 2014

[2] E V Christaki and P C Florou-Paneri ldquoAloe vera a plant formany usesrdquo Journal of Food Agriculture and Environment vol8 no 2 pp 245ndash249 2010

[3] S Rajasekaran K Ravi K Sivagnanam and S SubramanianldquoBeneficial effects of Aloe vera leaf gel extract on lipid profilestatus in rats with streptozotocin diabetesrdquo Clinical and Experi-mental Pharmacology and Physiology vol 33 no 3 pp 232ndash2372006

[4] S Shamim S W Ahmed and I Azhar ldquoAntifungal activity ofAlliumAloe and Solanum speciesrdquo Pharmaceutical Biology vol42 no 7 pp 491ndash498 2004

[5] M M Bottenberg G C Wall R L Harvey and S Habib ldquoOralaloe vera-induced hepatitisrdquo Annals of Pharmacotherapy vol41 no 10 pp 1740ndash1743 2007

[6] L Langmead RM Feakins S Goldthorpe et al ldquoRandomizeddouble-blind placebo-controlled trial of oral aloe vera gel foractive ulcerative colitisrdquoAlimentary Pharmacology ampTherapeu-tics vol 19 no 7 pp 739ndash747 2004

[7] S Li T Chen F Xu et al ldquoThe beneficial effect of exopolysac-charides from Bifidobacterium bifidum WBIN03 on microbialdiversity in mouse intestinerdquo Journal of the Science of Food andAgriculture vol 94 no 2 pp 256ndash264 2014

[8] T Chen S Li and H Wei ldquoAntibiotic resistance capability ofcultured human colonic microbiota growing in a chemostat

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Mediators of Inflammation 5

Emodin

lowastlowast

1 50 1025

Concentration (120583gmL)

05

06

07

08O

D570

(a)

Polysaccharide

30 501525 50

Concentration (120583gmL)

05

06

07

08

09

OD

570

(b)

Aloe veraAloe vera + L plantarum

05

06

07

08

09

OD

570

50 15 5030 200100

Concentration (120583gmL)

(c)

Figure 3 Different concentration of emodin (a) polysaccharide (b) Aloe vera and Aloe vera + L plantarum (c) on the cell viability of RAW2647 cells using MTT method Data are shown as the mean plusmn SD lowastlowast119875 lt 001

4 Discussion

Aswe all know the single target drug had been widely used invarious human diseases but often failed to cure the metabolicdiseases [28]Thus the whole-body systems biology was pro-posed and regarded the human and symbiotic microorgan-isms as a whole ldquosuperorganismrdquo [28 29] which led a moreand more reorganization by the scientific community to theChinese herb extracts and the human intestinal probiotics

Previously studies showed that both the AV [2 22] andprobiotics [25 30 31] had sound antioxidant and anti-inflammatory effects so we guessed that the combination ofthe AV and probiotics should possess better antioxidativeantibacterial and anti-inflammatory effects First we opti-mized the optimum growth temperature of the selected pro-biotics and found that the probiotics could grew well in AVextract without any additives (Figure 1) However no antimi-crobial effects of AV and Aloe fermentation were observedwhich was inconsistent with the previous studies [2 32]

(Figure 2) We guessed that the lack of glucose the substratefor probiotics to produce antimicrobial production of lacticacid and acetic acid was the cause of the antimicrobial failurefor Aloe fermentation therefore we added 5 glucose to AVextract and found that the added glucose not only signif-icantly increased the probiotics growth (Figure 1) but alsoconferred the Aloe fermentation supernatant sound antibac-terial effects on all the tested pathogens (Figure 2)

After a series of evaluations the L plantarumHM2187491was chosen and this strain conferred the Aloe fermentationsupernatant strong scavenging capacities of the DPPH (86)O2

∙minus (85) ∙OH (76) Fe2+ chelation (82) and reducingpowers (2425mgL) (Table 1) Moreover the Aloe fermen-tation fermented by L plantarum HM2187491 could inhibitall the growth of tested pathogens especially for the P acnesATCC 11827 (IZD 27mm)Therefore the sound inhibition ofthe growth ofP acnesATCC 11827 (causative bacteria for acnevulgaris) and antioxidant activitymade theAloe fermentationsupernatant a potential ingredient for cosmetic

6 Mediators of Inflammation

IL-1

120573 m

RNA

GA

PDH

mRN

A

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(a)

TNF-

a mRN

AG

APD

H m

RNA

0

10

20

30

40

lowastlowast

lowastlowast

lowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)

IL-6

mRN

AG

APD

H m

RNA

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 4 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the expression ofinflammatory genes (IL-1120573 TNF-120572 and IL-6) in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

As we know excess levels of IL-1120573 IL-6 and TNF-120572induced by activated inflammatory cells (eg eosinophilsmacrophages mononuclear phagocytes neutrophils) willlead to the damage of cells and tissues which eventually causethe inflammation-associated diseases for example rheuma-toid arthritis chronic hepatitis and macrophages play animportant role in regulating several immunopathologicalconditions and inducing overexpression of the proinflamma-tory mediators of the inflammatory process [24 25] So theRAW2647 cells were used in the present study to evaluate theanti-inflammatory effect of Aloe fermentation supernatantfermented by L plantarumHM2187491

The results indicated that both the emodin (anticanceractivity) [33] and the polysaccharide (immunity enhance-ment and reduction of oxidative injury) showed weakeranti-inflammatory effects compared to AV extract and Aloefermentation supernatant in mRNA level and protein level(Figures 4 and 5) Interestingly though theAloe fermentationsupernatant possessed a strong antioxidant effect (Figure 2)

its anti-inflammatory effect was inferior to the AV extractin both gene level and protein level Maybe some anti-inflammatory ingredients in AV extract were consumed byL plantarum HM2187491 and the metabolites of this strainmight contribute to the inflammatory response of RAW2647 cells Moreover in vivo work is needed because of theshortcomings of cell model for example it could not modelthe interaction between intestinal cells and microbiota Aspotential probiotics the L plantarum HM2187491 couldproduce bacteriocins offering advantages in colonization andcompetition in the gastrointestinal tract [34] and endowedthe AV extract a strong antioxidative effect and antimicrobialeffect on pathogens (especially for P acnes) In addition theL plantarumHM2187491 could remove the bitter taste of AVand our human results (119899 = 15) indicated that the Aloe fer-mentation supernatant had shortened the acne cure time andhad a perfect whitening effect (data unshown)

In conclusion the Aloe fermentation supernatantrsquos soundantimicrobial effect and anti-inflammatory effect together

Mediators of Inflammation 7

IL-1120573(p

gm

L)

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

lowastlowast

lowastlowast

lowastlowast lowastlowast

0

10

20

30

40

50

(a)

TNF-120572

(pg

mL)

0

500

1000

1500

lowastlowastlowastlowast lowastlowast

lowastlowast

lowastlowastlowastlowast lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)IL-6

(pg

mL)

lowastlowast

lowastlowast

lowastlowast lowastlowast lowastlowastlowastlowast

0

50

100

150

200

250

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 5 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the protein yields of IL-1120573TNF-120572 and IL-6 in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

with its intestinal health promoting effect when L plantarumHM2187491 enters host intestines indicate its potential useas functional foods or cosmetics to guard human intestinalhealth delaying senescence and preventing chronic diseases

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

This work was supported by grants from the National BasicResearch Program of China (2013CB531103) and NationalNatural Science Foundation of China (nos 8150336431560264 91339113 81270202 and 21461015) and the grantsfrom Jiangxi Province (nos 20151BAB205001 GJJ14218KJLD14010 and 20153BCB23035)

References

[1] Y-W Kim Y-J Jeong A-Y Kim et al ldquoLactobacillus brevisstrains from fermented Aloe vera survive gastroduodenal envi-ronment and suppress common food borne enteropathogensrdquoPLoS ONE vol 9 no 3 Article ID e90866 2014

[2] E V Christaki and P C Florou-Paneri ldquoAloe vera a plant formany usesrdquo Journal of Food Agriculture and Environment vol8 no 2 pp 245ndash249 2010

[3] S Rajasekaran K Ravi K Sivagnanam and S SubramanianldquoBeneficial effects of Aloe vera leaf gel extract on lipid profilestatus in rats with streptozotocin diabetesrdquo Clinical and Experi-mental Pharmacology and Physiology vol 33 no 3 pp 232ndash2372006

[4] S Shamim S W Ahmed and I Azhar ldquoAntifungal activity ofAlliumAloe and Solanum speciesrdquo Pharmaceutical Biology vol42 no 7 pp 491ndash498 2004

[5] M M Bottenberg G C Wall R L Harvey and S Habib ldquoOralaloe vera-induced hepatitisrdquo Annals of Pharmacotherapy vol41 no 10 pp 1740ndash1743 2007

[6] L Langmead RM Feakins S Goldthorpe et al ldquoRandomizeddouble-blind placebo-controlled trial of oral aloe vera gel foractive ulcerative colitisrdquoAlimentary Pharmacology ampTherapeu-tics vol 19 no 7 pp 739ndash747 2004

[7] S Li T Chen F Xu et al ldquoThe beneficial effect of exopolysac-charides from Bifidobacterium bifidum WBIN03 on microbialdiversity in mouse intestinerdquo Journal of the Science of Food andAgriculture vol 94 no 2 pp 256ndash264 2014

[8] T Chen S Li and H Wei ldquoAntibiotic resistance capability ofcultured human colonic microbiota growing in a chemostat

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

6 Mediators of Inflammation

IL-1

120573 m

RNA

GA

PDH

mRN

A

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(a)

TNF-

a mRN

AG

APD

H m

RNA

0

10

20

30

40

lowastlowast

lowastlowast

lowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)

IL-6

mRN

AG

APD

H m

RNA

0

50

100

150

200

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 4 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the expression ofinflammatory genes (IL-1120573 TNF-120572 and IL-6) in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

As we know excess levels of IL-1120573 IL-6 and TNF-120572induced by activated inflammatory cells (eg eosinophilsmacrophages mononuclear phagocytes neutrophils) willlead to the damage of cells and tissues which eventually causethe inflammation-associated diseases for example rheuma-toid arthritis chronic hepatitis and macrophages play animportant role in regulating several immunopathologicalconditions and inducing overexpression of the proinflamma-tory mediators of the inflammatory process [24 25] So theRAW2647 cells were used in the present study to evaluate theanti-inflammatory effect of Aloe fermentation supernatantfermented by L plantarumHM2187491

The results indicated that both the emodin (anticanceractivity) [33] and the polysaccharide (immunity enhance-ment and reduction of oxidative injury) showed weakeranti-inflammatory effects compared to AV extract and Aloefermentation supernatant in mRNA level and protein level(Figures 4 and 5) Interestingly though theAloe fermentationsupernatant possessed a strong antioxidant effect (Figure 2)

its anti-inflammatory effect was inferior to the AV extractin both gene level and protein level Maybe some anti-inflammatory ingredients in AV extract were consumed byL plantarum HM2187491 and the metabolites of this strainmight contribute to the inflammatory response of RAW2647 cells Moreover in vivo work is needed because of theshortcomings of cell model for example it could not modelthe interaction between intestinal cells and microbiota Aspotential probiotics the L plantarum HM2187491 couldproduce bacteriocins offering advantages in colonization andcompetition in the gastrointestinal tract [34] and endowedthe AV extract a strong antioxidative effect and antimicrobialeffect on pathogens (especially for P acnes) In addition theL plantarumHM2187491 could remove the bitter taste of AVand our human results (119899 = 15) indicated that the Aloe fer-mentation supernatant had shortened the acne cure time andhad a perfect whitening effect (data unshown)

In conclusion the Aloe fermentation supernatantrsquos soundantimicrobial effect and anti-inflammatory effect together

Mediators of Inflammation 7

IL-1120573(p

gm

L)

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

lowastlowast

lowastlowast

lowastlowast lowastlowast

0

10

20

30

40

50

(a)

TNF-120572

(pg

mL)

0

500

1000

1500

lowastlowastlowastlowast lowastlowast

lowastlowast

lowastlowastlowastlowast lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)IL-6

(pg

mL)

lowastlowast

lowastlowast

lowastlowast lowastlowast lowastlowastlowastlowast

0

50

100

150

200

250

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 5 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the protein yields of IL-1120573TNF-120572 and IL-6 in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

with its intestinal health promoting effect when L plantarumHM2187491 enters host intestines indicate its potential useas functional foods or cosmetics to guard human intestinalhealth delaying senescence and preventing chronic diseases

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

This work was supported by grants from the National BasicResearch Program of China (2013CB531103) and NationalNatural Science Foundation of China (nos 8150336431560264 91339113 81270202 and 21461015) and the grantsfrom Jiangxi Province (nos 20151BAB205001 GJJ14218KJLD14010 and 20153BCB23035)

References

[1] Y-W Kim Y-J Jeong A-Y Kim et al ldquoLactobacillus brevisstrains from fermented Aloe vera survive gastroduodenal envi-ronment and suppress common food borne enteropathogensrdquoPLoS ONE vol 9 no 3 Article ID e90866 2014

[2] E V Christaki and P C Florou-Paneri ldquoAloe vera a plant formany usesrdquo Journal of Food Agriculture and Environment vol8 no 2 pp 245ndash249 2010

[3] S Rajasekaran K Ravi K Sivagnanam and S SubramanianldquoBeneficial effects of Aloe vera leaf gel extract on lipid profilestatus in rats with streptozotocin diabetesrdquo Clinical and Experi-mental Pharmacology and Physiology vol 33 no 3 pp 232ndash2372006

[4] S Shamim S W Ahmed and I Azhar ldquoAntifungal activity ofAlliumAloe and Solanum speciesrdquo Pharmaceutical Biology vol42 no 7 pp 491ndash498 2004

[5] M M Bottenberg G C Wall R L Harvey and S Habib ldquoOralaloe vera-induced hepatitisrdquo Annals of Pharmacotherapy vol41 no 10 pp 1740ndash1743 2007

[6] L Langmead RM Feakins S Goldthorpe et al ldquoRandomizeddouble-blind placebo-controlled trial of oral aloe vera gel foractive ulcerative colitisrdquoAlimentary Pharmacology ampTherapeu-tics vol 19 no 7 pp 739ndash747 2004

[7] S Li T Chen F Xu et al ldquoThe beneficial effect of exopolysac-charides from Bifidobacterium bifidum WBIN03 on microbialdiversity in mouse intestinerdquo Journal of the Science of Food andAgriculture vol 94 no 2 pp 256ndash264 2014

[8] T Chen S Li and H Wei ldquoAntibiotic resistance capability ofcultured human colonic microbiota growing in a chemostat

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Mediators of Inflammation 7

IL-1120573(p

gm

L)

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

lowastlowast

lowastlowast

lowastlowast lowastlowast

0

10

20

30

40

50

(a)

TNF-120572

(pg

mL)

0

500

1000

1500

lowastlowastlowastlowast lowastlowast

lowastlowast

lowastlowastlowastlowast lowastlowast

lowastlowast

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(b)IL-6

(pg

mL)

lowastlowast

lowastlowast

lowastlowast lowastlowast lowastlowastlowastlowast

0

50

100

150

200

250

LPS

+50120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+5120583

gm

LAl

oe fe

rmen

tatio

n

LPS

+50120583

gm

LAl

oe ve

ra

LPS

+5120583

gm

LAl

oe ve

ra

LPS

+15

120583g

mL

poly

sacc

harid

e

LPS

+25120583

gm

Lpo

lysa

ccha

ride

LPS

+10

120583g

mL

emod

in

LPS

+25120583

gm

Lem

odin

LPS

Con

trol

(c)

Figure 5 Effect of the different concentrations of emodin polysaccharide Aloe vera and fermentation broth on the protein yields of IL-1120573TNF-120572 and IL-6 in RAW 2647 cells Data are shown as the mean plusmn SD lowastlowast119875 lt 001

with its intestinal health promoting effect when L plantarumHM2187491 enters host intestines indicate its potential useas functional foods or cosmetics to guard human intestinalhealth delaying senescence and preventing chronic diseases

Competing Interests

The authors declare that they have no competing interests

Acknowledgments

This work was supported by grants from the National BasicResearch Program of China (2013CB531103) and NationalNatural Science Foundation of China (nos 8150336431560264 91339113 81270202 and 21461015) and the grantsfrom Jiangxi Province (nos 20151BAB205001 GJJ14218KJLD14010 and 20153BCB23035)

References

[1] Y-W Kim Y-J Jeong A-Y Kim et al ldquoLactobacillus brevisstrains from fermented Aloe vera survive gastroduodenal envi-ronment and suppress common food borne enteropathogensrdquoPLoS ONE vol 9 no 3 Article ID e90866 2014

[2] E V Christaki and P C Florou-Paneri ldquoAloe vera a plant formany usesrdquo Journal of Food Agriculture and Environment vol8 no 2 pp 245ndash249 2010

[3] S Rajasekaran K Ravi K Sivagnanam and S SubramanianldquoBeneficial effects of Aloe vera leaf gel extract on lipid profilestatus in rats with streptozotocin diabetesrdquo Clinical and Experi-mental Pharmacology and Physiology vol 33 no 3 pp 232ndash2372006

[4] S Shamim S W Ahmed and I Azhar ldquoAntifungal activity ofAlliumAloe and Solanum speciesrdquo Pharmaceutical Biology vol42 no 7 pp 491ndash498 2004

[5] M M Bottenberg G C Wall R L Harvey and S Habib ldquoOralaloe vera-induced hepatitisrdquo Annals of Pharmacotherapy vol41 no 10 pp 1740ndash1743 2007

[6] L Langmead RM Feakins S Goldthorpe et al ldquoRandomizeddouble-blind placebo-controlled trial of oral aloe vera gel foractive ulcerative colitisrdquoAlimentary Pharmacology ampTherapeu-tics vol 19 no 7 pp 739ndash747 2004

[7] S Li T Chen F Xu et al ldquoThe beneficial effect of exopolysac-charides from Bifidobacterium bifidum WBIN03 on microbialdiversity in mouse intestinerdquo Journal of the Science of Food andAgriculture vol 94 no 2 pp 256ndash264 2014

[8] T Chen S Li and H Wei ldquoAntibiotic resistance capability ofcultured human colonic microbiota growing in a chemostat

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

8 Mediators of Inflammation

modelrdquo Applied Biochemistry and Biotechnology vol 173 no 3pp 765ndash774 2014

[9] M Kumar S Rakesh R Nagpal et al ldquoProbiotic Lactobacillusrhamnosus GG and Aloe vera gel improve lipid profiles inhypercholesterolemic ratsrdquoNutrition vol 29 no 3 pp 574ndash5792013

[10] A T Diplock P J Aggett M Ashwell F Bornet E B Fernand M B Roberfroid ldquoScientific concepts of functional foodsin Europe consensus documentrdquo British Journal of Nutritionvol 81 no 1 1999

[11] C Hill F Guarner G Reid et al ldquoExpert consensus documentthe International Scientific Association for Probiotics andPrebiotics consensus statement on the scope and appropriateuse of the term probioticrdquo Nature Reviews Gastroenterology ampHepatology vol 11 no 8 pp 506ndash514 2014

[12] D Granato G F Branco F Nazzaro A G Cruz and J A FFaria ldquoFunctional foods and nondairy probiotic food develop-ment trends concepts and productsrdquo Comprehensive Reviewsin Food Science and Food Safety vol 9 no 3 pp 292ndash302 2010

[13] E Parente F Ciocia A Ricciardi T Zotta G E Felis andS Torriani ldquoDiversity of stress tolerance in Lactobacillusplantarum Lactobacillus pentosus and Lactobacillus paraplan-tarum AMultivariate Screening Studyrdquo International Journal ofFood Microbiology vol 144 no 2 pp 270ndash279 2010

[14] W P Hammes and R F Vogel ldquoThe genus lactobacillusrdquo inTheGenera of Lactic Acid Bacteria pp 19ndash54 Springer BerlinGermany 1995

[15] X Wang Q Wu K Deng et al ldquoA novel method for screeningof potential probiotics for high adhesion capabilityrdquo Journal ofDairy Science vol 98 no 7 pp 4310ndash4317 2015

[16] H J M Harmsen G R Gibson P Elfferich et al ldquoComparisonof viable cell counts and fluorescence in situ hybridization usingspecific rRNA-based probes for the quantification of humanfecal bacteriardquo FEMS Microbiology Letters vol 183 no 1 pp125ndash129 2000

[17] T Chen Q Wu S Li et al ldquoMicrobiological quality and char-acteristics of probiotic products in Chinardquo Journal of the Scienceof Food and Agriculture vol 94 no 1 pp 131ndash138 2014

[18] L-S Lai S-T Chou and W-W Chao ldquoStudies on the antiox-idative activities of Hsian-tsao (Mesona procumbensHemsl) leafgumrdquo Journal of Agricultural and Food Chemistry vol 49 no 2pp 963ndash968 2001

[19] X Jiang X-C Huang L Ao et al ldquoTotal alkaloids of Triptery-gium hypoglaucum (levl) Hutch inhibits tumor growth both invitro and in vivordquo Journal of Ethnopharmacology vol 151 no 1pp 292ndash298 2014

[20] Z Yu C Jin M Xin and H JianMin ldquoEffect of Aloe verapolysaccharides on immunity and antioxidant activities in oralulcer animal modelsrdquo Carbohydrate Polymers vol 75 no 2 pp307ndash311 2009

[21] Y Hu J Xu and Q Hu ldquoEvaluation of antioxidant potentialof Aloe vera (Aloe barbadensis Miller) extractsrdquo Journal ofAgricultural and Food Chemistry vol 51 no 26 pp 7788ndash77912003

[22] D Babak and S N Nahashon ldquoA review on effects of aloe veraas a feed additive in broiler chicken dietsrdquo Annals of AnimalScience vol 14 no 3 pp 491ndash500 2014

[23] A McDowell S Valanne G Ramage et al ldquoPropionibacteriumacnes types I and II represent phylogenetically distinct groupsrdquoJournal of ClinicalMicrobiology vol 43 no 1 pp 326ndash334 2005

[24] D-J Yang Y-Y Chang H-W Lin Y-C Chen S-H Hsuand J-T Lin ldquoInhibitory effect of litchi (Litchi chinensis Sonn)Flower on lipopolysaccharide-induced expression of proinflam-matory mediators in RAW2647 cells through NF-120581B ERKand JAK2STAT3 inactivationrdquo Journal of Agricultural and FoodChemistry vol 62 no 15 pp 3458ndash3465 2014

[25] W-J Yoon Y M Ham S-S Kim et al ldquoSuppression ofpro-inflammatory cytokines iNOS and COX-2 expression bybrown algae Sargassum micracanthum in RAW 2647 macro-phagesrdquo EurAsian Journal of Biosciences vol 3 pp 130ndash1432009

[26] S Basannavar R Pothuraju and R K Sharma ldquoEffect of Aloevera (Aloe barbadensis Miller) on survivability extent of prote-olysis and ACE inhibition of potential probiotic cultures in fer-mentedmilkrdquo Journal of the Science of Food andAgriculture vol94 no 13 pp 2712ndash2717 2014

[27] C T Ramachandra and P S Rao ldquoProcessing of Aloe vera leafgel a reviewrdquo American Journal of Agricultural and BiologicalScience vol 3 no 2 pp 502ndash510 2008

[28] T Wang G Cai Y Qiu et al ldquoStructural segregation ofgut microbiota between colorectal cancer patients and healthyvolunteersrdquoThe ISME Journal vol 6 no 2 pp 320ndash329 2012

[29] F J Bruggeman and H V Westerhoff ldquoThe nature of systemsbiologyrdquo Trends in Microbiology vol 15 no 1 pp 45ndash50 2007

[30] M G Gareau P M Sherman and W A Walker ldquoProbioticsand the gut microbiota in intestinal health and diseaserdquo NatureReviews Gastroenterology amp Hepatology vol 7 no 9 pp 503ndash514 2010

[31] I P Kaur K Chopra and A Saini ldquoProbiotics potential phar-maceutical applicationsrdquo European Journal of PharmaceuticalSciences vol 15 no 1 pp 1ndash9 2002

[32] J Sutherland M Miles D Hedderley et al ldquoIn vitro effects offood extracts on selected probiotic and pathogenic bacteriardquoInternational Journal of Food Sciences and Nutrition vol 60 no8 pp 717ndash727 2009

[33] E Fenig J Nordenberg E Beery J Sulkes and L WassermanldquoCombined effect of aloe-emodin and chemotherapeutic agentson the proliferation of an adherent variant cell line ofMerkel cellcarcinomardquo Oncology Reports vol 11 no 1 pp 213ndash217 2004

[34] M P Castro N Z Palavecino C Herman O A Garro andC A Campos ldquoLactic acid bacteria isolated from artisanaldry sausages characterization of antibacterial compounds andstudy of the factors affecting bacteriocin productionrdquo MeatScience vol 87 no 4 pp 321ndash329 2011

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

MEDIATORSINFLAMMATION

of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Behavioural Neurology

EndocrinologyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Disease Markers

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

BioMed Research International

OncologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Oxidative Medicine and Cellular Longevity

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Computational and Mathematical Methods in Medicine

OphthalmologyJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Diabetes ResearchJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Research and TreatmentAIDS

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Gastroenterology Research and Practice

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom