research article phenolic acids (gallic and tannic acids...

Research ArticlePhenolic Acids (Gallic and Tannic Acids) Modulate AntioxidantStatus and Cisplatin Induced Nephrotoxicity in Rats

Seun F Akomolafe1 Ayodele J Akinyemi2 and Scholarstical O Anadozie2

1 Department of Biochemistry Ekiti State University PMB 5363 Ado Ekiti Nigeria2 Department of Biochemistry Afe Babalola University PMB 5454 Ado Ekiti Nigeria

Correspondence should be addressed to Seun F Akomolafe purposefulseunyahoocouk

Received 21 March 2014 Revised 21 May 2014 Accepted 2 June 2014 Published 11 August 2014

Academic Editor Ottavio Giampietro

Copyright copy 2014 Seun F Akomolafe et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Cisplatin (cis-diamminedichloroplatinum (II) or CDDP) used in the treatment of many solid-tissue cancers has its chief side-effect in nephrotoxicity Hence this study sought to investigate and compare the protective effect of gallic acid (GA) and tannicacid (TA) against cisplatin induced nephrotoxicity in rats The rats were given a prophylactic treatment of GA and TA orally at adose of 20 and 40mgkg body weight for 7 consecutive days before the administration of a single intraperitoneal (ip) injectionof cisplatin (CP) at 75mgkg bwt The protective effects of both GA and TA on CP induced nephrotoxicity were investigated byassaying renal function oxidative stress biomarkers and histopathological examination of kidney architecture A single dose ofcisplatin (75mgkg bwt) injected ip caused a significant increase in some biomarkers of renal function (creatinine uric acid andurea levels) with a marked elevation in malondialdehyde (MDA) content accompanied by a significant (119875 lt 005) decrease inreduced glutathione (GSH) content (10327) of kidney tissue as compared to control group Furthermore a significant (119875 lt 005)reduction in kidney antioxidant enzymes (SOD catalase GPx and GST) activity was observed However pretreatment with oraladministration of tannic acid and gallic acid at a dose of 20 and 40mgkg body weight respectively for 7 days prior to cisplatinadministration reduced histological renal damage and suppressed the generation of ROS lipid peroxidation and oxidative stress inkidney tissues These results indicate that both gallic and tannic acids could serve as a preventive strategy against cisplatin inducednephrotoxicity

1 Introduction

The use of chemotherapy in the treatment of cancer hasopened newpossibilities for improvement of the quality of lifeof cancer patients Despite its success treatment with some ofthe most effective anticancer drugs shows a number of symp-toms of direct toxicity [1] In recent years the mechanismof cisplatin (cis-diamminedichloroplatinum (II) or CDDP)induced nephrotoxicity has gradually been elucidated [2]Studies have shown an increase in lipid peroxides in therenal tissue of CDDP-administered animals [3] a decreasein reduced glutathione levels [1] and the induction ofmetallothionein [4] an antioxidantThese changes have beenconsidered to result from the generation of reactive oxygenspecies (ROS) Studies using chemiluminescence or electron

spin resonance (ESR) have shown that CDDP generates OHradical [5 6]

Nephrotoxicity involves kidney damage or dysfunctionarising from direct or indirect exposure to drugs andindustrial or environmental chemicals Cisplatin ((cis-diamminedichloroplatinum (II) or CDDP)) an anti-neoplastic drug have been reported to induce nephrotoxicity[7]The kidney which is themajor route of cisplatin excretionalso accumulates it to a greater degree than other organs[6 8] Oxidative stress inflammation and apoptosis are someof the mechanisms already established to explain cisplatininduced acute kidney injury [9] A number of strategies havebeen proposed for the preventionmanagement of cisplatininduced nephrotoxicity since there is no specific treatmentwith the use of some synthetic drugs which have been

Hindawi Publishing CorporationInternational Scholarly Research NoticesVolume 2014 Article ID 984709 8 pageshttpdxdoiorg1011552014984709

2 International Scholarly Research Notices

90

80

70

60

50

40

30

20

10

0

Gro

up1

Gro

up2

Gro

up3

Gro

up4

Gro

up5

Gro

up6

Groups

Nor

mal

cont

rol

Indu

ced

grou

p

MD

A le

vel (

nmol

g p

rote

in)

PlasmaKidney

lowast

and

and

andand

ampamp

amp amp

Figure 1 Effect of gallic and tannic acids on kidney and plasmaMDA contents in normal and cisplatin induced nephrotoxic rats 1Normal control 2 Induced control 3 20mg gallic acid + cisplatin4 40mg gallic acid + cisplatin 5 40mg gallic acid only 6 20mgtannic acid + cisplatin 7 40mg tannic acid + cisplatin and 840mg tannic acid only lowastSignificantly different (119875 lt 005) fromthe normal control ampandSignificantly (119875 lt 005) different from theinduced control

Gro

up1

Gro

up2

Gro

up3

Gro

up4

Gro

up5

Gro

up6

Groups

Nor

mal

cont

rol

Indu

ced

grou

p

10

9

8

7

6

5

4

3

2

1

0

lowast

Arg

inas

e act

ivity

(120583m

ol o

f ure

a pro

duce

dm

ing

pro

tein

)

Figure 2 Effect of gallic and tannic acids on arginase activity in thekidney of normal and cisplatin induced nephrotoxic rats 1 Normalcontrol 2 Induced control 3 20mg gallic acid + cisplatin 4 40mggallic acid + cisplatin 5 40mg gallic acid only 6 20mg tannic acid +cisplatin 7 40mg tannic acid + cisplatin and 8 40mg tannic acidonly lowastSignificantly (119875 lt 005) different from the normal controlSignificantly (119875 lt 005) different from the induced control

popular However these drugs have some associated risksand side-effects [9] hence the need for natural alternatives ofplant origin (plant foodsextracts) with little or no side-effect

Plants have limitless ability to synthesize aromatic sub-stances such as polyphenols mainly flavonoids and phenolicacids which exhibit antioxidant properties due to theirhydrogen-donating and metal-chelating capacities Polyphe-nols are secondary metabolites of plants and are widelydistributed in plant-derived foods such as cereals legumes

(a)

(b)

Figure 3 Histopathological views (times400) of the kidney showingsevere and generalized tubular epithelial cell necrosis associatedwith diffuse tubular lumina in rat administered a single ip dose ofcisplatin (75mgkg bwt) (b) while no damage is noticed in normalcontrol rats (a)

nuts vegetables and fruits and in beverages such as greenor black tea and fruit juice Several hundreds of differentpolyphenols have been identified in foods [10]

Tannic and gallic acids are two commonly phenolicacids that are structurally related Tannic acid a naturallyoccurring plant polyphenol is composed of a central glucosemolecule derivatized at its hydroxyl groups with one or moregalloyl residues whereas gallic acid is a trihydroxybenzoicacid also known as 345-trihydroxybenzoic acid which iswidely distributed in green tea red wine and grapes witchhazel sumac oak bark and other plants [11] Considerableamounts of experimental data on the antioxidant activity ofboth tannic acid and gallic acids with emphasis on structure-function antioxidant activity have been reported [12] Alsoseveral authors have demonstrated that tannic acid andother polyphenols have antimutagenic and anticarcinogenicactivities [13ndash16] Extensive studies have been carried outon the protective effect of cotreatment and posttreatment ofphenolic acids against cisplatin induced nephrotoxicity [17]Hence this study was carried out to investigate and comparethe protective effect of administration of both tannic andgallic acids on normal and cisplatin induced nephrotoxicityin rats

International Scholarly Research Notices 3

(a) (b)

(c) (d)

Figure 4 Histopathological views (times400) of the kidney showing a marked improvement on kidney damage in rats pretreated with gallic andtannic acids ((a) and (b)) gallic acid (20 and 40mgkg bwt) + cisplatin and ((c) and (d)) tannic acid (20 and 40mgkg bwt) + cisplatin

(a) (b)

Figure 5 Histopathological views (times400) of the kidney showing no damage in rats pretreated with gallic acid (40mgkg bwt) (a) and tannicacid (40mgkg bwt) (b) only without cisplatin

2 Materials and Methods

21 Experimental Animals Male albino rats weighing 110ndash185 g used for this experiment were purchased from a pri-vate animal colony Ikere-Ekiti metropolis The rats weremaintained at 25∘C on a 12 hour lightdark cycle with freeaccess to food and waterThey were acclimatized under theseconditions for two weeks prior to the commencement of theexperiments The experimental study was approved by theInstitutional Animal Ethical Committee of the University ofAdo-Ekiti Nigeria

22 Chemicals and Reagents Chemicals such as tannic acidgallic acid Oxidized and reduced glutathione hydrogen per-oxide (H

2O2) dithionitrobenzene (DTNB) thiobarbituric

acid (TBA) and adrenaline were purchased from SigmaChemical Co (St Louis MO USA) Ethanol acetic acidH2SO4 sodium carbonate sodium citrate sodium azide

(NaN3) sodium chloride potassium dichromate Tris-HCl

buffer sodium dodecyl sulphate (SDS) and Ascorbic acidwere sourced from BDH Chemicals Ltd (Poole England)Pharmaceutical grade cisplatin (CP) under the brand nameldquoCytoplatin 50rdquo was purchased from Cipla Ltd India May-Grunwald Giemsa and hematoxylin and eosin (HampE) stainswere purchased from Hi-Media Labs Mumbai All the kitsused for bioassay were sourced from RANDOX Laboratories


Table 1 Effect of tannic and gallic acids on renal function test in normal and cisplatin induced nephrotoxic rats

Groups Treatments Creatinine (mgdl) Urea (mgdl) UA (mgdl)1 Normal control 4404 plusmn 12 682 plusmn 08 712 plusmn 192 Induced control 13149b plusmn 23 851b plusmn 07 1109b plusmn 223 20mg gallic acid + cisplatin 4723c plusmn 18 705c plusmn 10 581c plusmn 084 40mg gallic acid + cisplatin 2043c plusmn 27 629c plusmn 15 496c plusmn 095 40mg gallic acid only 1659 plusmn 31 604 plusmn 16 480 plusmn 146 20mg tannic acid + cisplatin 6766c plusmn 23 731c plusmn 05 824c plusmn 357 40mg tannic acid + cisplatin 1149c plusmn 17 704c plusmn 05 433c plusmn 078 40mg tannic acid only 766 plusmn 04 653 plusmn 04 537 plusmn 13Values represent mean plusmn standard deviation (119899 = 6) bSignificantly (119875 lt 005) different from the normal control cSignificantly (119875 lt 005) different from theinduced control

Table 2 Effect of tannic acid and gallic acids on antioxidant content level in normal and cisplatin induced nephrotoxicity in rats

Groups SOD CAT GPx GSH1 201 plusmn 095 018 plusmn 010 021 plusmn 003 024 plusmn 0022 106b plusmn 053 009b plusmn 006 013b plusmn 000 013b plusmn 0053 131c plusmn 062 016c plusmn 003 017c plusmn 003 017c plusmn 0044 175c plusmn 124 016c plusmn 001 022c plusmn 005 018c plusmn 0025 205 plusmn 073 019 plusmn 002 023 plusmn 008 025 plusmn 0026 197c plusmn 093 012c plusmn 003 016c plusmn 000 017c plusmn 0037 239c plusmn 021 017c plusmn 020 023c plusmn 011 019c plusmn 0008 402 plusmn 095 018 plusmn 000 025 plusmn 014 022 plusmn 003Values represent mean plusmn standard deviation (119899 = 6) 1 induced control 2 normal control 3 20mg gallic acid + cisplatin 4 40mg gallic acid + cisplatin 540mg gallic acid only 6 20mg tannic acid + cisplatin 7 40mg tannic acid + cisplatin and 8 40mg tannic acid onlybSignificantly (119875 lt 005) different from the induced control cSignificantly (119875 lt 005) different from the induced control

Ltd Crumlin County Antrim UK Except stated otherwiseall other chemicals and reagents were of analytical grades andthe water was glass-distilled

23 Study Design and Treatment After two (2) weeks ofacclimatization 80 male rats were randomly divided intoeight (8) groups of ten animals each Group I served as anormal control and received saline (085 wv) orally for7 consecutive days and on the 7th day 1 hr after receivingthe oral saline dose the rats received a single ip injectionof saline (085) Group II served as toxicant group andreceived saline (085) orally for 7 consecutive days GAwas orally administered at two doses 20 and 40mgkgbody weight (bwt) to Groups III and IV respectively for 7consecutive days Also TA was orally administered at twodoses 20 and 40mgkg body weight (bwt) to Groups VI andVII respectively for 7 consecutive days On the 7th day ofpretreatment a single ip dose of cisplatin (75mgkg bwt)after oral treatment of GA and TA was given to the animalsin Groups II III IV VI and VII Groups V and VIII receivedonly higher dose (40mgkg) of GA and TA orally for 7consecutive days respectively and on the 7th day 1 hr GA andTA treatment received a single ip injection of saline (085)to ensure that Groups V and VIII received only the higherdose of GA and TA this was done in order to test that thehigher dose did not produce any kind of toxic effects All theanimals were killed after 24 hr of intoxication with cisplatinThe time of killing was based on the preliminary studies

The doses of TA and GA used were actually selected onthe basis of preliminary dose escalation studies to determinethe minimum dose of TA and GA required to produce anobservable effect (data not shown)

24 Sample Preparation All the animals were killed after24 hr of intoxication with cisplatin The animals were decap-itated after an overnight-fast by cervical dislocation Theblood was rapidly collected by direct heart puncture andthe plasma was prepared Uric acid urea and creatininewere determined using commercially available kits (Ran-dox Laboratories UK) Arginase activity was determinedas described by Kaysen and Strecker [18] Tissue malondi-aldehyde (MDA) content was determined as described byOhkawa et al [19] Tissue antioxidant parameters were alsodetermined superoxide dismutase (SOD) by the method ofAlia et al [20] catalase (CAT) by the method of Sinha [21]reduced glutathione (GSH) by the method of Ellman [22]and glutathione peroxidase (GPx) by the method of Rotrucket al [23]

25 Preparation of Plasma At the end of the experimentwhole blood of the sacrificed rats were collected into EDTAbottles and centrifuged at 800timesg for 10min to separate theplasma The plasma was then decanted into plain samplebottle and stored in a refrigerator prior to analysis


26 Preparation of Tissue Homogenates The ratrsquos tissues(kidney) were rapidly isolated placed on ice and weighedThe tissue was rinsed in cold (09wv) normal saline (1 3wv) and subsequently homogenized in sodium phosphatebuffer (pH 74) with (1 5 bt wv) using mortar and pestleas homogenizer and the homogenates were centrifuged at4000timesg The clear supernatants obtained were used forvarious biochemical assays [24]

27 Determination of Plasma Uric Acid Concentration Theuric acid concentration was determined using colorimetricmethod as described by Collin and Diehl [25] and Morinand Prox [26] Briefly 20120583L of distilled water was addedto 20120583L of the sample which was mixed with 1mL ofHepes reagent (50mM phosphate buffer 4mM 35-chloro-2-hydroxybenzenesulfonic acid) and enzyme reagent (025mM4-aminophenazone peroxidise and uricase) Thereafter themixture was incubated for 5min at 37∘C and the absorbanceat 520 nmwas taken against reagent blank within 30minTheuric acid concentration was subsequently calculated againstthe standard

28 Determination of Plasma Urea Concentration The ureaconcentration was determined using colorimetric methodas described by Searcy et al [27] Briefly 10 120583L of samplewas added to 01mL of sodiumnitroprussidemdashurease reagent(116mM EDTA 6mM sodium nitroprusside 1 gL urease)after which the mixture was incubated for 10min at 37∘C25mL of 120mM diluted phenol and 25mL of 27mMsodium hypochlorite solution containing 014N sodiumhydroxide which was then added to the reaction mixtureThereafter the mixture was incubated for 15min at 37∘C andthe absorbance at 546 nm was taken against reagent blankwithin 8 hours The urea concentration was subsequentlycalculated against the standard

29 Determination of Plasma Creatinine Concentration Thecreatinine concentration was determined using colorimetricalkaline picrate method as described by Jaffe method [28]Briefly 50 120583L of distilled water was added to 2mL of workingreagent (35mM picric acid and 032M sodium hydroxide)before 50 120583L of sample was added Thereafter the mixturewas allowed to stay for 30 seconds before taking absorbanceThe absorbance at 492 nm was taken twice firstly after 30 secand secondly after 2min The creatinine concentration wassubsequently calculated against the standard using change inthe sample absorbance (ΔAbsorbance)

210 Determination of Arginase Activity Arginase activitywas determined by the measurement of urea produced bythe reaction of Ehrlichrsquos reagent according to the modifiedmethod of Kaysen and Strecker [18] The reaction mixturecontained 10Mm Tris-HCL buffer 10mM MnCl

2(pH 95)

01M arginase solution and 500mL of the enzyme prepa-ration The mixture was incubated for 10mins at 37∘C Thereaction was terminated by the addition of 25mL Ehrlichreagent (20 g of p-dimethylaminobenzaldehyde in 200mL

of concentrated hydrochloric acid and made up to 100mLwith distilled water)

211 Determination of Superoxide Dismutase (SOD) ActivitySuperoxide dismutase (SOD) was determined by the methodof Alia et al [20] 50 120583L of supernatant was treated with1000 120583L of 50mM carbonate buffer (pH 102) and 17 120583L ofadrenaline (006mgmL)The absorbancewas read at 480 nmin spectrophotometer for 2 minutes at 15-second intervalsSOD activity was expressed as UI per 100 g protein (120576

480=

402mMminus1 cmminus1)

212 Determination of Catalase (CAT) Activity The activityof catalase (CAT) was determined by the method of Sinha[21]The reactionmixture (15mL) contained 10mLof 001Mphosphate buffer (pH 70) 01mL of tissue homogenateand 04mL of 2M H

2O2 The reaction was stopped by the

addition of 20mL of dichromate-acetic acid reagent (5potassium dichromate and glacial acetic acid were mixed in1 3 ratio) Then the absorbance was read at 620 nm CATactivity was expressed as moles of H

2O2consumedming

protein

213 Determination of Plasma Reduced Glutathione (GSH)Content Reduced glutathione (GSH) was determinedby the method of Ellman [22] 1mL of supernatantwas treated with 500120583L of Ellmanrsquos reagent (198mg of551015840dithiobisnitrobenzoic acid in 100mL of 01 sodiumcitrate) and 30mL of 02M phosphate buffer (pH 80) Theabsorbance was read at 412 nm in spectrophotometer

214 Determination of GPx Activity The activity of glu-tathione peroxidase (GPx) was assayed by the method ofRotruck et al [23]The reactionmixture containing 02mL ofEDTA (08mM Ph 70) 04mL of phosphate buffer (10mM)and 02mL of tissue homogenate was incubated with 01Mof H2O2and 02mL of glutathione for 10min Oxidation of

glutathione by the enzyme was measured spectrophotomet-rically at 420 nmThe activity of GPx was expressed as l120583molglutathione oxidizedming protein

215 Determination of Tissue Lipid Peroxidation The lipidperoxidation assay was carried out using the modifiedmethod of Ohkawa et al [19] Briefly 300 120583L of tissuehomogenate 300120583L of 81 SDS (sodium dodecyl sulphate)500120583Lof acetic acidHCl (pH=34) andTBA (thiobarbituricacid) were added and the mixture was incubated at 100∘Cfor 1 hr Thereafter the thiobarbituric acid reactive species(TBARS) produced was measured at 532 nm and calculatedas malondialdehyde (MDA) equivalent

216 Data Analysis The results of replicate readings werepooled and expressed as mean plusmn standard deviation One-way analysis of variance was used to analyze the results andDuncan multiple tests were used for the post hoc analysis[29] Statistical package for Social Science (SPSS) 100 for


Windows was used for the analysis The IC50

was calculatedusing nonlinear regression analysis

3 Results

As evident from Table 1 administration of a single doseof cisplatin (75mgkg bwt) caused a significant increase inthe biomarkers of renal function (creatinine urea and uricacid) when compared with the control group that receivedonly saline (085wv) However pretreatment with gallicand tannic acids orally at two doses 20 and 40mgkgbody weight respectively shows a significant (119875 lt 005)improvement of renal function due to a decrease in thecreatinine urea and uric acid levels when compared with theinduced group (Group 2)

Also administration of a single ip dose of cisplatin(75mgkg bwt) caused a significant decrease in the biomark-ers of oxidative stress [superoxide dismutase (SOD) catalase(CAT) glutathione peroxidase (GPx) and reduced glu-tathione (GSH)] when compared with the control group thatreceived only saline (085wv) (Table 2) However pretreat-ment with gallic and tannic acids orally at two doses 20and 40mgkg body weight respectively shows a significant(119875 lt 005) improvement in the bodyrsquos antioxidant status byan increase in the activities of superoxide dismutase (SOD)catalase (CAT) glutathione peroxidase (GPx) and reducedglutathione (GSH) when compared with the induced group(Table 2) Likewise therewas a significant (119875 lt 005) increasein the malondialdehyde (MDA) content in rat kidney admin-istered a single ip dose of cisplatin (75mgkg bwt) Howeverboth pretreatment with gallic and tannic acids orally at twodoses 20 and 40mgkg body weight respectively shows asignificant (119875 lt 005) reduction ofMDAcontent in rat kidneywhen compared with the induced group (Figure 1)

Figure 2 revealed that administration of a single ip doseof cisplatin (75mgkg bwt) caused a significant (119875 lt 005)increase in arginase activity when compared with the controlgroup that received only saline (085wv) However pre-treatment with gallic and tannic acids orally at two doses 20and 40mgkg body weight respectively shows a significant(119875 lt 005) decrease in arginase activity when compared withthe induced group (Group 2)

Photomicrographs of kidney sections from various treat-ment groups are shown in Figures 3 4 and 5 Histopatho-logical examination of sections from rat kidney administereda single ip dose of cisplatin (75mgkg bwt) shows severeand generalized tubular epithelial cell necrosis associatedwith diffuse tubular lumina when compared with the controlwithout cisplatin However pretreatment with gallic andtannic acids orally at two doses 20 and 40mgkg bodyweight respectively shows amarked improvement on kidneydamage

4 Discussion

About 25 of most commonly used drugs in intensive careunits (ICUs) are potentially nephrotoxic and are recognizedas considerable health and economic burden worldwide [1]

Among them cisplatin when used in cancer chemotherapyinduces renal impairment and acute renal failure by induc-tion of reactive oxygen species tubulointerstitial inflam-mation and apoptosis [7] Although various studies havebeen reported on the benefits of several agents in cisplatininduced renal toxicity the basis of nephroprotection remainselusive [6] This makes the search for strategies to preventnephrotoxicity constitute an active area of investigationAbundant evidences suggest the involvement of oxidativestress in the pathogenesis of cisplatin nephrotoxicity [8]Hence it is reasonable to assume that a reinforcement ofantioxidant defense of renal tissue by exogenous antioxidantsuch as phenolic acids should be a strategy to protect thekidney from the oxidative damage

In the present study we compare the protective effectof administration of gallic and tannic acids (two commonlyphenolic acids that are structurally related) of the samedosage against normal and cisplatin induced renal injury inrats for the first time by attenuating renal oxidative stressTheelevations of key kidney function biomarkers such as creati-nine uric acid and urea have been suggested to be indicativeof reduced renal functions [30 31]Thus estimation of plasmacreatinine and uric acid has been employed as key test toassess kidney function [30 31]

In the present study the observed elevation in plasmacreatinine urea and uric acid levels in induced rats indicatesa reduction in kidney function and hence nephrotoxicity(Table 1) This finding is consistent with that reported byearlier studies [32 33] However the restoration of the plasmacreatinine urea and uric acid level in rats treated withboth gallic and tannic acids (Table 1) suggests that bothphenolic acids have the ability to prevent kidney damageand protect the kidney against nephrotoxicity This howevermay be a function of their antioxidant properties and abilityto inhibit arginase activity (Figure 2) Cisplatin increasedarginase activity in the ratrsquos kidney while the pretreatmentwith both gallic and tannic acids at two doses (20 and40mgkg bwt) respectively for 7 days resulted in a decreasein kidney arginase activity (Figure 2)

Arginase is a hydrolytic enzyme responsible for theconversion of L-arginine to L-ornithine and urea Ornithineis an important biosynthetic precursor of polyamines whichhave been implicated to facilitate cell proliferation in certaincancer cells [34] Another important enzyme endotheliumnitric oxide synthase (eNOS) competes with arginase for thesame substrate arginine eNOS is involved in the productionof nitric oxide (NO) from arginine The NO produced playsan important role in both regulating renal hemodynamicsand modulating inflammatory and proliferating responseto various stimuli Therefore inhibition of arginase activityslows the progression of renal failure in renal ablation

The increase in the kidney and plasma MDA content(Figure 1) in the induced rats suggests lipid peroxidationThisagreed with earlier studies where administration of cisplatincaused inflammation and lipid peroxidation [6 7]This couldbe as a result of increased hydrogen peroxide concentrationproduced in the kidney due to the depletion of antioxidantenzymes superoxide dismutase (SOD) reduced glutathione(GSH) and catalase (CAT) activity (Table 2) This also is


consistent with earlier study where depletion of SOD CATandGPx in rats resulted in increasedMDAconcentration dueto lipid peroxidation [35]The depletion of these antioxidantssuggests that cisplatin induced nephrotoxicity could be aresult of oxidative stress or suppression of the antioxidantenzymes as previously reported by earlier studies [35ndash37]However the reduced kidney and plasma MDA content(Figure 1) and restoration of SOD GSH and CAT activities(Table 2) in pretreated rats suggest an improvement in thein vivo antioxidant status which may be a function of theantioxidant properties of the phenolic acids (gallic and tannicacids)

Reduced glutathione (GSH) has a multiple role as anantioxidant agent It functions as a scavenger of ROS includ-ing hydroxyl radicals and singlet oxygen [38] Thereforethe observed decrease in the kidney GSH level (Table 2) inthe induced rats suggests cisplatin induced nephrotoxicitywhich is associated with a drastic reduction in kidney GSHcontent This finding is consistent with earlier studies whereGSH depletion was suggested to be due to the interactionof cisplatin with the molecules contain sulfhydryl groups[39 40] However restoration of GSH levels in the pretreatedrats suggests the antioxidant and nephroprotective propertiesof the phenolic acids (Table 2)

Furthermore histopathology study revealed normalglomerulus and tubules with intact renal architecture innormal (Figure 3(a)) gallic (Figure 5(a)) and tannic acid(Figure 5(b)) group without cisplatin injection Degeneratedtubular structures with vacuolization and loss of architecturewere seen in cisplatin induced group (Figure 3(b)) Pretreat-ment with both gallic and tannic acids at two doses (20 and40mgkg bwt) respectively for 7 days resulted in excellentprotection against nephrotoxicity induced by cisplatin andshowed predominant normal kidney morphology (Figures4(a) 4(b) 4(c) and 4(d))

5 Conclusion

The results of the present study revealed that oxidative stressand apoptosisnecrosis play an important role in pathogen-esis of cisplatin nephrotoxicity Gallic and tannic acids twoimportant pharmacologically active phenolic compoundsreduced cisplatin induced functional and histological renaldamage Furthermore they suppressed the generation ofROS lipid peroxidation and oxidative stress in kidney tis-sues These results indicated that both gallic and tannic acidsexhibit nephroprotective effect and the possible mechanismof action by which they exert this effect could be dueto their antioxidant properties and inhibition of arginaseactivity However tannic acid exhibited better nephropro-tective potential than gallic acid which may be due to theglycosidation with a glucose moiety

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] E M El-Sayed M F Abd-Ellah and S M Attia ldquoProtectiveeffect of captopril against cisplatin-induced nephrotoxicity inratsrdquo Pakistan Journal of Pharmaceutical Sciences vol 21 no 3pp 255ndash261 2008

[2] R S Goldstein and G H Mayor ldquoThe nephrotoxicity ofcisplatin minireviewrdquo Life Sciences vol 32 no 7 pp 685ndash6901983

[3] M Satoh N Kashihara S Fujimoto et al ldquoA novel free radicalscavenger edarabone protects against cisplatin-induced acuterenal damage in vitro and in vivordquoThe Journal of Pharmacologyand Experimental Therapeutics vol 305 no 3 pp 1183ndash11902003

[4] P J Boogaard E L Lempers G J Mulder and J H NMeerman ldquo4-Methylthiobenzoic acid reduces cisplatin nephro-toxicity in rats without compromising anti-tumour activityrdquoBiochemical Pharmacology vol 41 no 12 pp 1997ndash2003 1991

[5] H Masuda M Fukumoto K Hirayoshi and K NagataldquoCoexpression of the collagen-binding stress protein HSP47gene and the alpha 1(I) and alpha 1(III) collagen genes in carbontetrachloride-induced rat liver fibrosisrdquo The Journal of ClinicalInvestigation vol 94 no 6 pp 2481ndash2488 1994

[6] R P Miller R K Tadagavadi G Ramesh and W B ReevesldquoMechanisms of cisplatin nephrotoxicityrdquo Toxins vol 2 no 11pp 2490ndash2518 2010

[7] A Sreedevi K Bharathi and K V S R G Prasad ldquoEffect ofdecoction of root bark of Berberis aristata against cisplatin-induced nephrotoxicity in ratsrdquo International Journal of Phar-macy and Pharmaceutical Sciences vol 2 no 3 pp 51ndash56 2010

[8] R K Tadagavadi and W B Reeves ldquoEndogenous IL-10 atten-uates cisplatin nephrotoxicity role of the total antioxidantcapacity and the activity of liver antioxidant enzymes in ratsrdquoNutrional Resources vol 23 pp 1251ndash1267 2010

[9] G Dong J Luo V Kumar and Z Dong Inhibitors of His-tone Deacetylases Suppress Cisplatin-Edition Clarendon PressOxford UK 2010

[10] A Scalbert andGWilliamson ldquoDietary intake andbioavailabil-ity of polyphenolsrdquo Journal of Nutrition vol 130 supplement 8pp 2073Sndash2085S 2000

[11] L D Reynolds and N G Wilson Scribes and Scholars OxfordUniversity Press Oxford UK 3rd edition 1991

[12] C Rice-Evans N J Miller and G Paganga ldquoStructure-antioxidant activity relationships of flavonoids and phenolicacidsrdquo Free Radical Biology andMedicine vol 20 no 7 pp 933ndash956 1996

[13] Y Nakamura A Kaihara K Yoshii Y Tsumura S Ishimitsuand Y Tonogai ldquoEffects of the oral administration of green teapolyphenol and tannic acid on serum and hepatic lipid contentsand fecal steroid excretion in ratsrdquo Journal of Health Science vol47 no 2 pp 107ndash117 2001

[14] T Okuda T Yoshida and T Hatano ldquoHydrolyzable tanninsand related polyphenolsrdquo Progress in the Chemistry of OrganicNatural Products vol 66 pp 1ndash115 1995

[15] A E Hagerman KM Riedl G A Jones et al ldquoHighmolecularweight plant polyphenolics (tannins) as biological antioxidantsrdquoJournal of Agricultural and Food Chemistry vol 46 no 5 pp1887ndash1892 1998

[16] MM Cowan ldquoPlant products as antimicrobial agentsrdquoClinicalMicrobiology Reviews vol 12 no 4 pp 564ndash582 1999

[17] K Tikoo D K Bhatt A B Gaikwad V Sharma and D GKabra ldquoDifferential effects of tannic acid on cisplatin induced


nephrotoxicity in ratsrdquo The FEBS Letters vol 581 no 10 pp2027ndash2035 2007

[18] G A Kaysen and H J Strecker ldquoPurification and properties ofarginase of rat kidneyrdquo Biochemical Journal vol 133 no 4 pp779ndash788 1973

[19] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[20] M Alia C Horcajo L Bravo and L Goya ldquoEffect of grapeantioxidant dietary fibre on antioxidant system and causesoxidation in rat kidney tissues possibole protective roles ofnatural antioxidant foodsrdquo Journal of Applied Toxicology vol 26pp 42ndash46 2003

[21] A K Sinha ldquoColorimetric assay of catalaserdquo Analytical Bio-chemistry vol 47 no 2 pp 389ndash394 1972

[22] G L Ellman ldquoTissue sulfhydryl groupsrdquo Archives of Biochem-istry and Biophysics vol 82 no 1 pp 70ndash77 1959

[23] J T Rotruck A L Pope H E Ganther A B Swanson D GHafeman and W G Hoekstra ldquoSelenium biochemical role asa component of glatathione peroxidaserdquo Science vol 179 no4073 pp 588ndash590 1973

[24] N A V Belle G D Dalmolin G Fonini M A Rubin and JB T Rocha ldquoPolyamines reduces lipid peroxidation induced bydifferent pro-oxidant agentsrdquo Brain Research vol 1008 no 2pp 245ndash251 2004

[25] P F Collin and H Diehl ldquoDeterminationof uric acidrdquo JustusLiebigs Annalen der Chemie vol 31 pp 1862ndash1867 1959

[26] L G Morin and J Prox ldquoReduction of ferric phenanthroline aprocedure for determining serum uric acidrdquo American Journalof Clinical Pathology vol 60 no 5 pp 691ndash694 1973

[27] R L Searcy J E Reardon and J A Foreman ldquoA newphotometric method for serum urea nitrogen determinationrdquoThe American Journal of Medical Technology vol 33 no 1 pp15ndash20 1967

[28] F W Spierto M L MacNeil and C A Burtis ldquoThe effect oftemperature and wavelength on the measurement of creatininewith the Jaffe procedurerdquoClinical Biochemistry vol 12 no 1 pp18ndash21 1979

[29] J H Zar Biostatistical Analysis Prentice Hall Upper SaddleRiver NJ USA 1984

[30] W Arneson and J Brickell ldquoAssessment of renal functionrdquo inClinical Chemistry A Laboratory Perspective pp 201ndash232 F ADavis Philadelphia Pa USA 2007

[31] P B Godkar ldquoKidney function testsrdquo in Text Book of MedicalLaboratory Technology Bhalani Publishing House BombayIndia 1994

[32] H M F Abdel-Wahab N I Y Hassanin E M Ahmed andA R Abdel-Wahab ldquoImpact of dried black grape andor hotred pepper supplementation in ameliorating the nephrotoxicityeffect of cisplatin in ratsrdquoAustralian Journal of Basic andAppliedSciences vol 5 no 10 pp 231ndash238 2011

[33] M V Makwana N M Pandya D N Darji S A Desai and VH Bhaskar ldquoAssessment of nephroprotective potential of Sidacordifolia Linn in experimental animalsrdquoDer Pharmacia Lettrevol 4 no 1 pp 175ndash180 2012

[34] C W Tabor and H Tabor ldquoPolyaminesrdquo Annual Review ofBiochemistry vol 53 pp 749ndash790 1984

[35] MM Ahmed ldquoBiochemical studies on nephroprotective effectof carob (Ceratonia siliqua L) growing in Egyptrdquo Nature andScience vol 8 no 3 pp 41ndash47 2010

[36] T A Ajith N Jose and K K Janardhanan ldquoAmeliorationof cisplatin induced nephrotoxicity in mice by ethyl acetateextract of a polypore fungus Phellinus rimosusrdquo Journal ofExperimental and Clinical Cancer Research vol 21 no 2 pp213ndash217 2002

[37] R Cetin E Devrim B Kilicoglu A Avci I Candir andI Durak ldquoCisplatin impairs antioxidant system and causesoxidation in rat kidney tissues possible protective roles ofnatural antioxidant foodsrdquo Journal of Applied Toxicology vol 26no 1 pp 42ndash46 2006

[38] B Halliwell and JM C Gutteridge Free Radicals in Biology andMedicine Clarendon Press Oxford UK 2nd edition 1989

[39] E M El-sayed M F Abd-ellah and S M Attia ldquoProtectiveeffect of captopril against flavonoids and phenolic acidsrdquo FreeRadicals in Biology and Medicine vol 20 pp 933ndash956 2008

[40] M J Khoshnoud B N A Moghbel B Geramizadeh and HNiknaha ldquoEffect of simvastatin on cisplatin-induced nephro-toxicity inmale ratsrdquo Iranian Journal of Pharmaceutical Sciencesvol 7 no 3 pp 165ndash173 2011

Submit your manuscripts athttpwwwhindawicom

PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

ToxinsJournal of

VaccinesJournal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

AntibioticsInternational Journal of

ToxicologyJournal of


StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Drug DeliveryJournal of



Advances in Pharmacological Sciences

Tropical MedicineJournal of


Medicinal ChemistryInternational Journal of


AddictionJournal of



BioMed Research International

Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Autoimmune Diseases


Anesthesiology Research and Practice

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of


Pharmaceutics


MEDIATORSINFLAMMATION

of


90

80

70

60

50

40

30

20

10

0

Gro

up1

Gro

up2

Gro

up3

Gro

up4

Gro

up5

Gro

up6

Groups

Nor

mal

cont

rol

Indu

ced

grou

p

MD

A le

vel (

nmol

g p

rote

in)

PlasmaKidney

lowast

and

and

andand

ampamp

amp amp

Figure 1 Effect of gallic and tannic acids on kidney and plasmaMDA contents in normal and cisplatin induced nephrotoxic rats 1Normal control 2 Induced control 3 20mg gallic acid + cisplatin4 40mg gallic acid + cisplatin 5 40mg gallic acid only 6 20mgtannic acid + cisplatin 7 40mg tannic acid + cisplatin and 840mg tannic acid only lowastSignificantly different (119875 lt 005) fromthe normal control ampandSignificantly (119875 lt 005) different from theinduced control

Gro

up1

Gro

up2

Gro

up3

Gro

up4

Gro

up5

Gro

up6

Groups

Nor

mal

cont

rol

Indu

ced

grou

p

10

9

8

7

6

5

4

3

2

1

0

lowast

Arg

inas

e act

ivity

(120583m

ol o

f ure

a pro

duce

dm

ing

pro

tein

)

Figure 2 Effect of gallic and tannic acids on arginase activity in thekidney of normal and cisplatin induced nephrotoxic rats 1 Normalcontrol 2 Induced control 3 20mg gallic acid + cisplatin 4 40mggallic acid + cisplatin 5 40mg gallic acid only 6 20mg tannic acid +cisplatin 7 40mg tannic acid + cisplatin and 8 40mg tannic acidonly lowastSignificantly (119875 lt 005) different from the normal controlSignificantly (119875 lt 005) different from the induced control

popular However these drugs have some associated risksand side-effects [9] hence the need for natural alternatives ofplant origin (plant foodsextracts) with little or no side-effect

Plants have limitless ability to synthesize aromatic sub-stances such as polyphenols mainly flavonoids and phenolicacids which exhibit antioxidant properties due to theirhydrogen-donating and metal-chelating capacities Polyphe-nols are secondary metabolites of plants and are widelydistributed in plant-derived foods such as cereals legumes

(a)

(b)

Figure 3 Histopathological views (times400) of the kidney showingsevere and generalized tubular epithelial cell necrosis associatedwith diffuse tubular lumina in rat administered a single ip dose ofcisplatin (75mgkg bwt) (b) while no damage is noticed in normalcontrol rats (a)

nuts vegetables and fruits and in beverages such as greenor black tea and fruit juice Several hundreds of differentpolyphenols have been identified in foods [10]

Tannic and gallic acids are two commonly phenolicacids that are structurally related Tannic acid a naturallyoccurring plant polyphenol is composed of a central glucosemolecule derivatized at its hydroxyl groups with one or moregalloyl residues whereas gallic acid is a trihydroxybenzoicacid also known as 345-trihydroxybenzoic acid which iswidely distributed in green tea red wine and grapes witchhazel sumac oak bark and other plants [11] Considerableamounts of experimental data on the antioxidant activity ofboth tannic acid and gallic acids with emphasis on structure-function antioxidant activity have been reported [12] Alsoseveral authors have demonstrated that tannic acid andother polyphenols have antimutagenic and anticarcinogenicactivities [13ndash16] Extensive studies have been carried outon the protective effect of cotreatment and posttreatment ofphenolic acids against cisplatin induced nephrotoxicity [17]Hence this study was carried out to investigate and comparethe protective effect of administration of both tannic andgallic acids on normal and cisplatin induced nephrotoxicityin rats


(a) (b)

(c) (d)


(a) (b)

























2(pH 95)




480=





2O2consumedming

protein









3 Results





4 Discussion











5 Conclusion




References















































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


(a) (b)

(c) (d)


(a) (b)

























2(pH 95)




480=





2O2consumedming

protein









3 Results





4 Discussion











5 Conclusion




References















































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

















2(pH 95)




480=





2O2consumedming

protein









3 Results





4 Discussion











5 Conclusion




References















































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of




3 Results





4 Discussion











5 Conclusion




References















































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of





5 Conclusion




References















































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of






























Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

research article phenolic acids (gallic and tannic acids...

Documents