pleurotus ostreatus produces antioxidant and anti

www.wjpps.com Vol 4, Issue 05, 2015.

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Singh et al. World Journal of Pharmacy and Pharmaceutical Sciences

PLEUROTUS OSTREATUS PRODUCES ANTIOXIDANT AND ANTI-

ARTHRITIC ACTIVITY IN WISTAR ALBINO RATS

Vinita Singh2*

, Dr. Deepak Vyas1, Prof. Rajshree Pandey

2 and Imtiyaz Ahmad Sheikh

1

1Lab of Microbial Technology, Dept. of Botany, Dr. H. S. Gour (Central) University, Sagar-

470003 (M.P.) India

2Dept. of Botany, A.P.S. University, Rewa (M.P.) India.

ABSTRACT

Pleurotus ostreatus is a distinguished cultivable oyster mushroom with

remarkable mycoremediation and mycotherapeutic properties. The aim

of this study was to evaluate the therapeutic claims of aqueous extract

of P. ostreatus (AEPO) in relieving arthritic conditions. In this study the

aqueous extract of P. ostreatus was also screen to display potent

antioxidant activity in vitro, total phenolic and flavonoid contents in

order to find possible sources for future novel antioxidants in food and

pharmaceutical formulations. A detailed study was performed on the

antioxidant activity of aqueous extract of P. ostreatus by DPPH

scavenging method and formalin induced arthritic injuries, lipid peroxidation in

wistar-albino rats. The total phenolic content (TPC) and total flavonoid content (TFC) of the

extract were also determined. Paw thickness, White Blood Cell Count (WBC), Hemoglobin

Concentration (HB), Erythrocyte Sedimentation Rate (ESR), lipid peroxidation, catalase

activity were studied post induction of arthritis. The extracts of P. ostreatus were also

subjected to preliminary phytochemical screening test for various constituents. The total

phenolic contents (119.9±3.1 mg GAE/g extract) and total flavonoid contents (60.9 ±2.2 mg

QE/g extract) of aqueous extract of P. ostreatus (AEPO) were found to be significantly high.

The IC50 value of AEPO on the DPPH radical was 42.0±1 μg/ml. Results of in-vivo

experiment revealed that administration of formalin induced arthritis injury caused a

significant increase in lipid peroxidation compare to normal saline group. In contrast, AEPO

(300 mg/kg bw) and standard drug (Dexamethasone) (50 mg/kg bw) treatments effectively

prevented these alterations and maintained the antioxidant status. Mean paw thickness in

animals given 300 mg kg1 AEPO was significantly (p<0.05) lower on days 7 and 14 compared

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VVoolluummee 44,, IIssssuuee 0055,, 11223300--11224466.. RReesseeaarrcchh AArrttiiccllee IISSSSNN 2278 – 4357

Article Received on

06 March 2015,

Revised on 27 March 2015,

Accepted on 18 April 2015

*Correspondence for

Author

Vinita Singh

Dept. of Botany, A.P.S.

University, Rewa (M.P.)

India.

http://www.wjpps.com/


1231


to that of normal saline group. Data from present results revealed that P. ostreatus act as an

antioxidant agent due to its free radical scavenging and exhibited potent anti-arthritic activity.

KEY WORDS: Pleurotus ostreatus, formalin, arthritis, oxidative stress.

1. INTRODUCTION

Pleurotus ostreatus, the oyster mushroom, is a common edible mushroom grown

commercially around the world and widely used for their high nutritional value as a

functional food. Oyster mushrooms can also be used industrially

for mycoremediation purposes. The oyster mushroom may be considered as medicinal

mushroom, since it contains statins such as lovastatin which work to reduce cholesterol (Eger

et al., 1976). Free radicals are basic requirement to any biochemical process and represent an

essential part of aerobic life and metabolism (Tiwari et al., 2001). Reactive oxygen specice

have been implicated in over a hundred of diseases which range from arthritis and connective

tissue disorders to carcinogenesis, aging, physical injury, infection and acquired

immunodeficiency syndrome (Joyce, 1987). Phenolic compounds widely distributed in

mushrooms which have been reported to exert multiple biological effects including

antioxidant, free radical scavenging abilities, anti-inflammatory, anti-carcinogenic etc

(Miller, 1996). The anti oxidative and free radical scavenging properties of the phenolic

content of mushroom aqueous extracts have been reported, suggesting possible protective

roles of these compounds, due to their ability to capture metals, inhibit lipoxygenase and

scavenge free radicals (Mau, Chang, Huang, & Chen, 2004). Recently, (Valentao et al., 2005)

identified the presence of six phenolic compounds (3-, 4- and 5-O-caffeoylquinic acid, caffeic

acid, p-coumaric acid and rutin) and five organic acids (citric, ascorbic, malic, shikimic and

fumaric acids) in edible mushrooms. Joint diseases are mainly classified as inflammatory or

non inflammatory disorders (Halliwell & Gorman, 1989). Degenerative joint disease

(osteoarthritis) is a non-inflammatory joint disorder while inflammatory diseases of the joint

include feline progressive polyarthritis, lupus polyarthritis, idiopathic non erosive arthritis

and rheumatoid arthritis a broad range in auto immune diseases (Todhunter & Johnston,

2003; Hegen et al., 2008). This joint disorder also affects other organs and tissue such as the

heart, lungs, eyes, kidney and neuromuscular system (Mahajan et al., 2010). Rheumatoid

Arthritis (RA) is a chronic auto immune-mediated disease which affects humans and animals

(Kaur et al., 2012). In the joint, RA is characterized by profuse inflammatory reaction in the

synovial membrane and subchondral bone which results in progressive erosion of articular


http://en.wikipedia.org/wiki/Mycoremediation

http://en.wikipedia.org/wiki/Medicinal_mushroom

http://en.wikipedia.org/wiki/Medicinal_mushroom

http://en.wikipedia.org/wiki/Statins

http://en.wikipedia.org/wiki/Lovastatin

http://en.wikipedia.org/wiki/Cholesterol


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cartilage and synovitis (Hegen et al., 2008; Kaur et al., 2012). In advanced cases, ankylosis,

subluxation, soft tissue destruction, disuse osteoporosis and pain may be noticed (Halliwell,

& Gorman, 1989; Goldring & Goldring, 2006).

There etiology for RA is unknown but several drugs such as anti-inflammatory and disease

modifying anti-rheumatoid drugs are used in mono or combination therapies to inhibit the

disease process (Makinen et al., 2007; Zhao et al., 2006; Mottonen et al., 2006). However,

prolonged use of these drugs is associated with deleterious side effects such as gastric

ulceration, haemorrhage, anemia and kidney dysfunction (Lin et al., 2006; Buhroo, & Baba,

2006; Kyei et al., 2012). Thus in recent times, researches have been directed towards the use

of biologics and plant-derived drugs in the treatment of RA (Woode et al., 2009; Kaithwas,

& Majumdar, 2010). Fresh mushrooms contain significant levels of l-ergothioneine, which

acts as an antioxidant. Beta-glucans, a type of carbohydrate found in mushrooms, has

potential anti-inflammatory activity, which may help protect the body against disease.

Mushroom extract may also stimulate different cells of the immune system. P. sajor caju

extract exhibit a strong anti-inflammatory effect, potential protective effect and

immunomodulary effect mediated by immune mechanisms. (Patel et al.,2012).

2. MATERIAL AND METHODS

2.1. Preparation of Mushroom extract

Strains of P. ostreatus (MTCC 142) were grown at Lab of Microbial Technology, Dept. of

Botany, Dr. Hari Singh Gour Central University, Sagar (M.P.) India. The pure fungal Species

procured from IMTECH, (Chandigarh), India. The fruiting bodies were dried in an oven at

450C for 4 hours. The dried fruiting bodies were crushed to powder by using electronic

blender. About 50g of powder were taken in 500ml of distilled water in Soxhlet extraction

unit for extraction at 300C for 18-20 hours and filtered through Whatman No. 4 filter paper.

The extract was evaporated almost to dryness in a rotary evaporator (Rotavapor R-114,

Buchi) at 40 °C and then subjected to freeze drying (LYOVAC, GEA).

2.2. Preliminary Phytochemical Screening

The phytochemical screening of the aqueous extract of P.ostreatus (AEPO) was performed

according to standard literature methods in which the extract were exposed to different

reagents to identify the primary metabolites, like carbohydrates (Molisch reagent test),

amino acids(millions test) and the secondary metabolites such as alkaloids (Mayer’s test),

flavonoids, terpenoids (Salkowski test), tannins (Ferric chloride test).



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2.3. Scavenging effect on 1, 1-Diphenly-2- picrylhydrazyl (DPPH)

The hydrogen atom or electron donating ability of the AEPO was measured from the

bleaching of the purple-colored methanol solution of 1, 1- Diphenly-2-picrylhydrazyl

(DPPH). This spectrophotometric assay uses the stable radical DPPH as a reagent (Burits, M.,

& Bucar, F., 2000). 1000 µl of various concentrations (20-200µg/ml) of the extract were

added to 4 ml of 0.004% methanol solution of DPPH. After 20-30 min incubation period at

room temperature, the absorbance was read against a blank at λ = 517 nm. Inhibition of free

radical by DPPH in percent (%I) was calculated by the following formula.

% inhibition = (A blank – A sample / A blank) x 100

Where A blank is the absorbance of the control (Solvent) and A-sample is the absorbance of

the test compound.

Extract concentration providing 50% inhibition (IC50) was calculated from the graph plotted

inhibition percentage against extract concentration. Tests were carried out in triplicate.

2.4. Determination of total phenolic compounds (TPC)

Total soluble phenolic in the mushroom aqueous extract was determined with Folin-

Ciocalteu reagent according to the method of (Slinkard & Singleton, 1977) by using Gallic

acid as a standard. 1.0 ml mushroom extract solution was taken in a volumetric flask and was

diluted with 45ml of distilled water. 1ml Folin-Ciocalteu reagent was added and mixed

thoroughly. After 5 min, 2ml of Na2CO3 (2%) was added and the mixture was allowed to

stand for 2½ hours with intermittent shaking. The absorbance of developed blue colour was

measured at 760 nm. The concentration of total phenols was expressed as mg/g of the dry

extract (Kim et al., 2003). Absorbance = 0.0009 × gallic acid (μg)

2.5. Determination of total flavonoid contents (TFC)

The aluminum chloride colorimetric method was used to measure the flavonoid

content of all plant extract. Extract solution (0.25ml, 1mg/ml) of each plant extract

was added to 1.25 ml of distilled water. Sodium nitrite solution (0.075ml, 5%) was then

added to the mixture followed by incubation for 5 minutes after which 0.15ml of 10%

aluminium chloride was added. The mixture was allowed to stand for 6min at room

temperature before 0.5ml of 1 M sodium hydroxide was finally added and the mixture diluted

with 0.275 ml distilled water. The absorbance of the reaction mixture was measured at 510



1234


nm with a UV/VIS spectrophotometer immediately. Quercetin was used as the standard for

the calibration curve. Flavonoid contents were expressed as mg quercetin equivalent (QE)/g

dry weight (D.W.).

2.6. Experimental animals

Wistar Albino rats of either sex (weighing 100-150 g) were obtained from Department of

Research and Defense Organization (DRDO), Gwalior, (M.P.). The animals were maintained

in a well-ventilated room, fed on standard pellet feed and water ad libitum. All studies on

animals were approved by Institutional Animal Ethics Committee (Approved no. IAEC

/2013/13).

2.7. Formaldehyde-Induced Arthritis

Non-immunological arthritis was induced in four groups (n = 5) of rats by sub plantar

injection of 0.1 ml Freshly prepared 2.5% formaldehyde (Seyle, 1949) on day 1 and repeated

on day 3. Groups 1 and 2 received 100 and 300 mg kg1 body weight (b.w) (AEPO) orally

(p.o) while groups 3 and 4 received Dexamethasone (50 mg/kg bw) (i.m) and normal saline (1

mg kg1, p.o) 1 h before arthritis induction respectively. The paw thicknesses of rats were

measured on days 0, 3, 5, 7 and 10 using a venire caliper. The edema component of arthritis

was estimated by calculating the difference between day 0 paw thicknesses and paw

thicknesses at the various time points. Blood samples were collected from retro orbital plexus

on days 0, 3, 7 and 10 for total white blood cell count (Bain et al., 2012). On day 10, rats

were euthanized and paw tissues collected to determine Malondialdehyde equivalents (MDA)

level as well as Superoxide Dismutase (SOD) and Catalase (CAT) activities in rat paw

tissues. MDA equivalent was determined as described by (Ohkawa et al.1979). SOD activity

was estimated using the procedure of (Sun et al. 1988) while CAT was assayed as described

by (Sinha, 1972). MDA Equivalent, SOD and CAT levels in paw tissues of a non-

arthritic/normal group (group 5) were also studied and compared with those of the

arthritic/treatment groups. On day 28, blood was collected to determine the Hemoglobin

Concentrations (HB), red Blood Cell Counts (RBC), White Blood Cell counts (WBC) and

Erythrocyte Sedimentation Rates (ESR) of rats. HB was determined by the

cyanmethaemoglobin method, RBC and WBC were determined by the haemocytometer

method while ESR was determined using the Westergren method a described by (Bain et al.

2012). HB, RBC, WBC and ESR of non-arthritic/normal group (group 5) were also studied

and compared with those of the arthritic/treatment groups.



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2.8. Statistical Analysis

Data obtained were compared between groups using one way Analysis of Variance

(ANOVA). Duncan multiple range tests were used to test significant differences between

means at the level p<0.05.

3. RESULTS AND DISCUSSION

3.1. Preliminary Phytochemical Screening

Preliminary phytochemical tests revealed that the main active constituents of P. ostreatus

extract are polyphenols including flavonoids, tannins, amino acids, and simple phenols were

present.

3.2 Inhibition of DPPH radical

DPPH is a stable free radical at room temperature and accepts an electron or hydrogen radical

to become a stable diamagnetic molecule (Amar-owicz, Peg g, Mogh addam, Barl, & Weil,

2004). The reduction capability of DPPH was determined by the decrease in its absorbance at

λ=517 nm, which is induced by antioxidants. Positive DPPH test suggests that the aqueous

extract of the mushroom was free radical scavenger. The scavenging effect of mushroom

extract and ascorbic acid on DPPH radical was compared.100 µg /ml of mushroom extract

and ascorbic acid exhibited 77.99% and 107.87% inhibition respectively (figure.1). The IC50

values were found to be (59.07±2.01 μg/ml) and (42.0±01 μg/ml), for ascorbic acid and

mushroom, respectively. The different concentrations of mushroom extract (20, 40, 60, 80

and 100 µg /ml) showed antioxidant activities in a dose dependent manner on DPPH radical.

From the present results, it may be postulated that compounds present in AEPO reduced the

radical to the corresponding hydrazine when it reacted with hydrogen donors in the

antioxidant principles. DPPH radicals react with suitable reducing agents, the electrons

become paired off and the solution loses color stoichometrically depending on the number of

electrons taken up (Sanchez-Moreno, C. 2002). In the present study, it was observed that

purple color of DPPH was bleached completely and very rapidly by the extract at all

concentrations (20-200µg/ml) in a concentration dependent manner indicating very efficient

scavenging activity of AEPO.

Oxidative stress has been implicated in the pathology of many diseases, such as Parkinson

disease, Alzheimer, diabetes, cardio vascular disorders, aging and inflammatory conditions

etc. The results obtained in the present studies may be attributed to several reasons i.e. DPPH



1236


radicals, total phenolic and flavonoid content generated in the mammalian cells, involved in

the regulation of various physiological processes. However, excess production of these

radicals is associated with several diseases (Aneta et al,.1993).

3.3 Determination of (TPC) and (TFC)

The flavonoid content of the extract in terms of quercetin equivalent (Figure 2)shows in

Table 2 .Table 2 also shows the contents of total phenols that were measured by

Folin-Ciocalteu reagent in terms of gallic acid equivalent (Figure 3). In all, aqueous

extract generally, exhibited the higher values of antioxidants. The result clearly shows that

the solvent influences the extractability of the phenolic compounds. This confirms the

assertion that phenolic content of mushroom contribute directly to their antioxidant

properties. Which was found to be 119.9 μg/mg gallic acid equivalents of phenols .The 20

μg/mg of phenols in any mushroom is considered to be sufficient for antioxidant activity,

therefore, and our results are much better as it contains adequate proportion of phenols (119.9

μg/mg gallic acid equivalents of phenols). Phenols are basic plant constituents and deserve

important because of their scavenging ability due to their hydroxyl groups (Harborne, 1989).

(Duh et al., 1999) showed that the phenolic compounds in mushrooms may contribute

directly to antioxidative action.

Flavonoids are naturally occurring in mushroom and are thought to have possible and

encouraging effects on human health. Studies on flavonoidic derivatives have shown a wide

range of antibacterial, antiviral, anti inflammatory, anticancer and antiallergic activities (Di

Carlo G, Mascolo N, Izzo A.A., Capasso, F.1999), (Montoro, P. Braca A, Pizza C, De

Tommasi, N. 2005). Flavonoids have been shown to be highly effective scavengers of most

oxidizing molecules, including singlet oxygen, and various free radicals (Bravo, L., 1998).

3.4. Formaldehyde-Induced Arthritis

The effect of P.ostreatus extracts on formaldehyde induced arthritis is summarized in

Table.3, 4, 5, 6. The results showed that mean paw thickness of 300 and 100 mg kg1 (AEPO)

groups were significantly (p<0.05) lower than that of normal saline group. Mean WBC of 100

and 300 mg kg1 (AEPO) groups were significantly (p<0.05) lower than that of normal saline

group (Table4). Mean MDA equivalent levels of normal saline and 100 mg kg1 (AEPO)

groups were significantly (p<0.05) higher than MDA equivalent levels of non-arthritic and

300 mg kg1 (AEPO) groups. MDA Equivalent levels of 300 mg kg1 (AEPO) and

dexamethasone groups were similar to that of non-arthritic group (Table 5). Mean SOD



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levels of 300 and 100 mg kg1 (AEPO) groups were significantly (p<0.05) higher than SOD

level of normal saline group. Mean catalase level of 100 and 300 mg kg1 AEPO groups were

significantly (p<0.05) higher compared to that of normal saline group (Table 5).

The inflammatory process is a physiologic response of a living organism to factors such as

infection, trauma or immunological mechanisms (Tanas et al., 2010). This progression is

initiated by the host to eliminate irritants and to set the stage for tissue repair (Bhitre et al.,

2008). Formaldehyde injection elicits localized inflammation and pain in the early phase

followed subsequently by a phase of tissue mediated response (Aceto & Cowan, 1991). This

late phase produces proliferative joint inflammation leading to articular changes similar to

those seen in rheumatoid arthritis (Okoli et al., 2008). Arthritis induced by formalin is model

used for the estimate of an agent with probable antiproliferative activity (Banerjee et al.,

2000). Acute inflammation induced by formalin result from cell damage, which provokes

the creation of endogenous mediators, such as, histamine, serotonin, prostaglandins, and

bradykinin (Yuh-Fung et al., 1995).Thus, formaldehyde induced arthritis are commonly used

experimental models for preclinical screening of non-steroidal anti-inflammatory drugs, disease

modifying anti-rheumatoid drugs and plant extracts for anti-arthritic effect (Woode et al.,

2009). The results obtained in this study showed that (AEPO) significantly suppressed

formaldehyde induced arthritis as shown by the significantly lesser paw and joint thickness in

300 mg kg1 (AEPO) group post arthritis induction. However, measurement of paw and joint

thickness gives only an indication of edematous changes in these regions (Woode et al.,

2009); therefore to correlate the edematous changes with the local biochemical changes,

tissue MDA Equivalent, SOD and catalase level activities in rat paw were measured during

formaldehyde-induced arthritis.

Phagocytes such as macrophages and neutrophils which invade inflamed tissues generate

reactive oxygen species (Valko et al., 2006). ROS apart from being defensive, when in

excess deregulate cellular function causing oxidative damage which worsens inflammation

(Wu et al., 2006; Tanas et al., 2010). Cells contain a number of anti-oxidants such as

superoxide dismutase, catalase and glutathione peroxidase which prevent the damage caused

by ROS (Weydert and Cullen, 2010). SOD converts superoxide radical to hydrogen peroxide

and oxygen while catalase decomposes hydrogen peroxide into water (Weydert & Cullen,

2010). In this study, SOD and catalase levels of normal saline and 100 mg kg1 groups were

significantly lower than that of non-arthritic rats. Earlier studies have shown that SOD and



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catalase levels decreased in chronic inflammatory states (Halici et al., 2007; Wu et al., 2006;

Govindarajan et al., 2007). Thus we can conclude that the presence of formaldehyde in the

tissues stimulated a profuse production of ROS which significantly besieged the antioxidant

system in rat paw tissues leading to the decrease in SOD level. However, SOD and catalase

levels in the paws of rats treated with 300 mg kg1 (AEPO) were almost parallel to that of

non-arthritic rats suggesting that severe paw inflammation was attenuated by administration

of this treatment. The level of MDA Equivalent in the tissue is considered a measure of lipid

peroxidation which is linked to the production of superoxide radical (Karatas et al., 2003).

Increased level of MDA equivalent as seen in normal saline group was an indication that the

presence of formaldehyde in the tissues stimulated profuse production of free radicals.

Increased lipid peroxidation in rat paw tissues following injection of irritants such as

carrageenan has been reported (Tanas et al., 2010). Furthermore, the lower MDAequivalent

level in the 300 mg kg1 (AEPO) and dexamethasone group suggests that both treatments

ameliorated the inflammatory process thus dampening the production of free radicals. In

formaldehyde induced arthritis, WBC of the normal saline treated rats was higher than those

of the other treatment groups. The increase in WBC in all the groups followed the same

pattern as the degree of paw inflammation. Previously, leucocytosis and neutrophilia

characterized adjuvant-induced arthritis in rats (Franch et al., 1994). White blood cells are

chief components of the host defense system thus the increased WBC seen in this study can

be attributed to systemic response of the rats to paw inflammation induced by formaldehyde.

(Franch et al., 1994). Furthermore, lower WBC in the 300 mg kg1 (AEPO)group suggests

that the extract showed potent anti-arthritic effect given that elevated WBC are associated

with active inflammation (Kyei, 2012).

HB and RBC of normal saline and (AEPO) groups were not significantly different from

those of non-arthritic rats. Earlier, (Kyei, 2012) reported that RBC and HB of rats were not

affected post induction of arthritis. However, this author reported a significant raise in ESR

during arthritis. Our finding also showed that ESR was significantly faster in normal saline

and in 100 mg kg1 (AEPO)groups but was slower in 300 mg kg1 (AEPO)group. Therefore,

since proteins produced during inflammation cause erythrocytes to stack up in a group

leading to faster settling (Kyei, 2012), the altitude of ESR observed in 100 mg kg1 (AEPO)

group showed the presence of high quantity of inflammatory proteins in circulation while the

near normal ESR in 300 mg kg1 (AEPO) group points to the fact that inflammation was less

severe in this group, because ESR work as inflammatory marker.



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Table1. Preliminary Phytochemical screening of the various extracts of the P.ostreatus

Chemical Constituent Solvents

Aqueous Methanol Ethyl

acetate

Pet-

ether

Alkaloid 1.Dragendroff’s Reagent

2. Mayer’s Reagent

3. Wagner’s Reagent

4. Hager Reagent

-

+

+

+

-

+

+

+

-

+

+

-

-

+

-

-

Amino acid Millon’s Test - - - -

Carbohydrate 1.Molish Test + + + -

2. Barfoed’s Test + + + -

Flavonoid Sample + Lead acetate + + + -

Tanin Ferric Chloride Test + + - -

Figure-1 DPPH scavenging activity of P.ostreatus aqueous extract at different

concentrations

Table2. Total Phenolic and flavonoid contents of the P.ostreatus extracts

Table 3. Effect of P.ostreatus extract on changes in paw thickness during formaldehyde

induced arthritis {Mean paw thickness (mm)}

Groups Day 0 Day 3 Day 7 Day 14

Extract Phenolic Content (mg of GAE/g of extract) Flavonoid

content (mg of QE/g of extract).

Aqueous 119.9± 3.1 60.9 ± 2.2

Methanol 48.01± 2.0 33.16 ± 1.7

Ethyl acetate 35.22 ± 3.7 21.69 ± 7.0

Pet-ether 26.01 ± 1.3 14.26 ± 3.4



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1 0.32±0.01 0.59±0.01a 0.52±0.02

a 0.49±0.02

a

2 0.54±0.01 0.62±0.03a 0.56±0.03

a 0.51±0.02

a

3 0.44±0.01 0.65±0.02a 0.57±0.03

a 0.51±0.01

a

4 0.46±0.01 0.78±0.04b 0.84±0.02

b 0.78±0.06

b

Group 1 = formaldehyde +300 mg kg1AEPO; group 2 = formaldehyde +100 mg kg1; AEPO

group 3 = formaldehyde +50 mg kg1dexamethasone; group 4 = formaldehyde + normal

saline. Different superscriptsa,b

in a column show significant differences at level p<0.05

Table4. White blood cell counts in P.ostreatus and dexamethasone treated rats during

formaldehyde-induced arthritis {WBC (109/uL)}

Groups Day 0 Day 3 Day 7 Day 10

1 6.70±0.10 07.80±0.10a 07.00±0. 10b 6.80±0.20

b

2 6.20± 0.30 07.60±0.50a 08.70±0.30

c 8.50±0.30

c

3 6.90±0.50 06.90±0.90a 04.70±0.50

d 4.60±0.40

d

4 6.50±0.60 12.50±0. 10b 15.10±0.20

a 14.1±0.10

a




in a column show significant difference at level p<0.05

Table5. Malondialdehyde Equivalent (MDA), Superoxide Dismutase (SOD) and

Catalase (CAT) levels in paw tissues during formaldehyde-induced arthritis

Groups MDA (nmoL/g tissue) SOD (units/g tissue) Catalase (units/g tissue)

1 04.35±0.69a 214. 10±1 .50

c 20.60±2.22

c

2 18.50±2.99b 131.30±2.02

b 11.10±0.98

b

3 07.40±1 .23a 251.50±3.60

d 23.00±2.68

c

4 69.80±10.77c 085. 10±2.60

a 00.22±0.03

a

5 02.40±0.60a 261.40±3.90

d 27.40±2.43

d




in a column show significant differences at level p<0.05

Table6. Hematologic parameters of rats treated with P.ostreatus extract and

dexamethasone during formaldehyde-induced arthritis



saline. Group 5=Control group, Different superscripts a, b

in a column show significant

difference at level p<0.05

Supplementary file

Figure of P.ostreatus

Groups HB(g/dL) RBC(103/µL) WBC(10

9/µL) ESR (mm/hr)

1 12.97±0.20b 6.35±0.20

ab 11.05±1.28

a 2.50±1.00

a

2 12.40±0.20b 5.58±0.42

ab 15.00±1.90

a 4.50±0.50

b

3 9.30±1.50a 3.53±0.60

a 09.98±2.30

a 1.80±0.70

a

4 14.45±0.55b 4.87±0.14

ab 28.13±2.40

b 6.00±0.50

c

5 13.60±0.20b 6.79±0. 13

b 08.30±8.80

a 2.30±0.33

a



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Figure 4 Hematologic parameters of rats treated with P.ostreatus extract and

dexamethasone during formaldehyde-induced arthritis

4 . C ON CL USI ON

According to the several references which reported anti-inflammatory, antibacterial and

antitumoral activities of this mushroom have previously been investigated (Mau et al., 2004),

Based on a comprehensive report on the DPPH scavenging activity of Pleurotus ostreatus, it

has exhibited excellent antioxidant activity that could have significance as therapeutic

agents in preventing inflammation and ageing associated oxidative stress related

degenerative diseases. T h i s study showed that daily administration of 300 mg kg1

significantly ameliorated the arthritic progression as shown by lesser local (paw edema and

tissue anti-oxidant activities) and systemic (WBC and ESR) changes in rats treated with this

dose of AEPO. Hence we bring to a close that P.ostreatus can serve as a respectable anti-

arthritic agent and can be potential source of natural antioxidant compounds for pharmaceutical

application.

ACKNOWLEDGEMENT

Author is thankful to Head of the Department of Botany, Dr. H S Gour (Central) University,

Sagar, (MP) for providing lab facilities for continuity and successfully running research

work.

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