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RMACEUTIC
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Singh
Sciences
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Volume 4, Issue 05, 1230-1246.

Research Article

ISSN 2278 4357

PLEUROTUS OSTREATUS PRODUCES ANTIOXIDANT AND ANTIARTHRITIC ACTIVITY IN WISTAR ALBINO RATS
Vinita Singh2*, Dr. Deepak Vyas1, Prof. Rajshree Pandey2 and Imtiyaz Ahmad Sheikh1
1

Lab of Microbial Technology, Dept. of Botany, Dr. H. S. Gour (Central) University, Sagar470003 (M.P.) India
2

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

Article Received on
06 March 2015,

ABSTRACT

Revised on 27 March 2015,


Accepted on 18 April 2015

remarkable mycoremediation and mycotherapeutic properties. The aim

Pleurotus ostreatus is a distinguished cultivable oyster mushroom with

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


*Correspondence for

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

Author

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

Vinita Singh

antioxidant activity in vitro, total phenolic and flavonoid contents in

Dept. of Botany, A.P.S.

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

University, Rewa (M.P.)


India.

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 (119.93.1 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 42.01 g/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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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

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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 (Mayers 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 l of various concentrations (20-200g/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 = 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 FolinCiocalteu 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 2 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 (g)
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

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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 nonarthritic/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
=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 g /ml of mushroom extract
and ascorbic acid exhibited 77.99% and 107.87% inhibition respectively (figure.1). The IC50
values were found to be (59.072.01 g/ml) and (42.001 g/ml), for ascorbic acid and
mushroom, respectively. The different concentrations of mushroom extract (20, 40, 60, 80
and 100 g /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 (20-200g/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

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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 g/mg gallic acid equivalents of phenols .The 20
g/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
g/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
Petacetate
ether
1.Dragendroffs Reagent
Alkaloid
2. Mayers Reagent
+
+
+
+
3. Wagners Reagent
+
+
+
4. Hager Reagent
+
+
Amino acid
Millons Test
+
+
+
Carbohydrate 1.Molish Test
2. Barfoeds Test
+
+
+
Sample
+
Lead
acetate
+
+
+
Flavonoid
Ferric Chloride Test
+
+
Tanin

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


concentrations
Table2. Total Phenolic and flavonoid contents of the P.ostreatus extracts
Extract Phenolic Content (mg of GAE/g of extract)
Flavonoid
content (mg of QE/g of extract).
119.9 3.1
60.9 2.2
Aqueous
48.01 2.0
33.16 1.7
Methanol
35.22 3.7
21.69 7.0
Ethyl acetate
26.01 1.3
14.26 3.4
Pet-ether

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


induced arthritis {Mean paw thickness (mm)}
Groups
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1
0.320.01
0.590.01a
0.520.02a
0.490.02a
2
0.540.01
0.620.03a
0.560.03a
0.510.02a
a
a
3
0.440.01
0.650.02
0.570.03
0.510.01a
4
0.460.01
0.780.04b
0.840.02b
0.780.06b
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
6.700.10
07.800.10a
07.000. 10b
6.800.20b
2
6.20 0.30
07.600.50a
08.700.30c
8.500.30c
a
d
3
6.900.50
06.900.90
04.700.50
4.600.40d
b
a
4
6.500.60
12.500. 10
15.100.20
14.10.10a
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 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
04.350.69a
214. 101 .50c
20.602.22c
b
b
2
18.502.99
131.302.02
11.100.98b
3
07.401 .23a
251.503.60d
23.002.68c
4
69.8010.77c
085. 102.60a
00.220.03a
a
d
5
02.400.60
261.403.90
27.402.43d
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
Table6. Hematologic parameters of rats treated with P.ostreatus extract and
dexamethasone during formaldehyde-induced arthritis
Groups
HB(g/dL)
RBC(103/L)
WBC(109/L)
ESR (mm/hr)
1
12.970.20b
6.350.20ab
11.051.28a
2.501.00a
2
12.400.20b
5.580.42ab
15.001.90a
4.500.50b
a
a
a
3
9.301.50
3.530.60
09.982.30
1.800.70a
4
14.450.55b
4.870.14ab
28.132.40b
6.000.50c
b
b
a
5
13.600.20
6.790. 13
08.308.80
2.300.33a
Group 1 = formaldehyde +300 mg kg1AEPO; group 2 = formaldehyde +100 mg kg1; AEPO
group 3 = formaldehyde +50 mg kg1dexamethasone; group 4 = formaldehyde + normal
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

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


dexamethasone during formaldehyde-induced arthritis
4. CONCLUSION
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 antiarthritic 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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