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Antiparkinsonian Activity of Aqueous Extract of Agaricus Blazei Murill in Rotenone-induced Parkinson's Disease

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Background: Parkinsons disease is a chronic neurological disorder which may be due to reduction in the dopaminergic neurons in the brain. However, Agaricus blazei is a rich source of natural antioxidants. Aim: In this study, antiparkinsonian activity of Agaricus blazei Murill was evaluated using different animal models. Method: Antiparkinsonian activity was evaluated using two different doses (273 mg/kg and 819 mg/kg) of Agaricus blazei Murill. Rotenone and sunflower oil were used as positive and negative control, respectively. Catalepsy test, rotarod test, exploratory behavior test (rearing) and locomotor activity test were conducted to observe antiparkinsonian activity of the drug in rats. Result: The results of the animal models were confirmed by determining the levels of reduced glutathione, total protein, thiobarbituric acid reactive substances (TBARS) and nitric oxide in the animal brain. Pretreatment with Agaricus blazei Murill, showed marked reduction in rotenone-Original Research Article Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598 122 induced catalepsy and a significant increase in exploratory behavior, muscular activity, and locomotor activity in rats. Agaricus blazei Murill has also shown extremely significant effect in decreasing the oxidative stress in the animal brain by increasing the brain levels of reduced GSH and total proteins and decreasing the levels of nitrite and TBARS. Conclusion: The results of rotenone-induced catalepsy, exploratory behavior, rotarod test and locomotor activity showed that Agaricus blazei Murill exerts a significant ameliorative effect on Parkinson's disease in rats.
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*Corresponding author: E-mail: Pharmacologist1@yahoo.com;
Journal of Pharmaceutical Research International
33(33B): 121-131, 2021; Article no.JPRI.69598
ISSN: 2456-9119
(Past name: British Journal of Pharmaceutical Research, Past ISSN: 2231-2919,
NLM ID: 101631759)
Antiparkinsonian Activity of Aqueous Extract of
Agaricus Blazei Murill in Rotenone-induced
Parkinson’s Disease
Muhammad Aslam
1*
, Hammad Ahmed
2
, Tayyaba Mumtaz
2
and Gahzal Hakani
3
1
Department of Pharmacology, Faculty of Pharmacy, University of Sindh, Jamshoro 76080, Pakistan.
2
Department of Pharmacology, Faculty of Pharmacy, Ziauddin University, Karachi 75600, Pakistan.
3
Department of Pharmacognosy, Faculty of Pharmacy, Ziauddin University, Karachi 75600, Pakistan.
Authors’ contributions
This work was carried out in collaboration among all authors. Author MA designed the study, wrote
the protocol, and wrote the first draft of the manuscript. Author HA wrote rest of the manuscript and
managed the analyses of the study. Author TM and GH managed the final review of the manuscript.
All authors read and approved the final manuscript.
Article Information
DOI: 10.9734/JPRI/2021/v33i33B31803
Editor(s):
(1) Dr. Paola Angelini, University of Perugia, Italy.
Reviewers:
(1) Dong Hwan Ho, Wonkwang University, South Korea.
(2) Sathyamurthy. P, Sri Ramachandra University, India.
Complete Peer review History:
http://www.sdiarticle4.com/review-history/69598
Received 20 April 2021
Accepted 27 June 2021
Published 29 June 2021
ABSTRACT
Background:
Parkinsons disease is a chronic neurological disorder which may be due to reduction
in the dopaminergic neurons in the brain. However, Agaricus blazei is a rich source of natural
antioxidants.
Aim: In this study, antiparkinsonian activity of Agaricus blazei Murill was evaluated using different
animal models.
Method: Antiparkinsonian activity was evaluated using two different doses (273 mg/kg and 819
mg/kg) of Agaricus blazei Murill. Rotenone and sunflower oil were used as positive and negative
control, respectively. Catalepsy test, rotarod test, exploratory behavior test (rearing) and locomotor
activity test were conducted to observe antiparkinsonian activity of the drug in rats.
Result: The results of the animal models were confirmed by determining the levels of reduced
glutathione, total protein, thiobarbituric acid reactive substances (TBARS) and nitric oxide in the
animal brain. Pretreatment with Agaricus blazei Murill, showed marked reduction in rotenone-
Original Research Article
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
122
induced catalepsy and a significant increase in exploratory behavior, muscular activity, and
locomotor activity in rats. Agaricus blazei Murill has also shown extremely significant effect in
decreasing the oxidative stress in the animal brain by increasing the brain levels of reduced GSH
and total proteins and decreasing the levels of nitrite and TBARS.
Conclusion: The results of rotenone-induced catalepsy, exploratory behavior, rotarod test and
locomotor activity showed that Agaricus blazei Murill exerts a significant ameliorative effect on
Parkinson’s disease in rats.
Keywords: Agaricus blazei; antiparkinsonian; antioxidants; oxidative stress and Parkinson’s disease.
1. INTRODUCTION
Parkinson’s disease (PD) may be due to the
selective damage of dopaminergic neurons in
substantia nigra pars compacta. It has been
observed that oxidative stress plays a major role
in the pathophysiology of Parkinson’s Disease.
Reactive oxygen species (ROS) are formed due
to oxidative stress which result in neuronal death
of the neurons This can be detected by
decreased levels of endogenous antioxidants.
Therefore, the use of antioxidants, along with
other protective agents could be a better
therapeutic intervention in PD. The current
therapeutic agents, because of various side
effects, have failed to prove to be a cure-all
therapy for the PD patients [1]. Hence, the
search for safer alternative/complementary
medicines for the management of PD is an
unfinished task for the researchers in this area of
study.
Fungi, like primary basidiomycetes and
mushrooms, are one of the significant edible
foods. Numerous of these mushrooms are
important because of their ability to produce
some ingredients of high medicinal importance.
Such mushrooms are called medicinal
mushrooms [2]. A medicinal mushroom called
Agaricus blazei Murill (ABM) commonly known
as Cogumelo do sol, family Agariaceae, has a
rich medicinal history. Traditionally, ABM has
been well known for the management of physical
and emotional stress, improvement and
stimulation of immunity, and improvement in
diabetic conditions. The herb is also potentially
effective to fight against hypercholesterinemia,
osteoporosis, peptic ulcer and digestive
problems and different kinds of cancers
Orivasios and Apuleius used the herb for the
treatment of ulcers of the larynx in malignant
conditions [2,3]. Scientifically, ABM is known for
its pharmacological activities including,
anticancer activity [4], immunostimulant and
immunomodulatory activities [5,6], antiviral
activity [7], antibacterial activity [8],
cardioprotective effect [9], weight controlling and
hypolipidemic effect [10] and hepatoprotective
effect [11]. Antioxidants are famous for their
neuroprotective potential [12]. Agaricus blazei
Murill is an abundant source of naturally occuring
antioxidants and aqueous extract of ABM has
shown significant in vitro antioxidant activity [13].
This study was designed to investigate if
Agaricus blazei possesses antiparkinsonian
activity due to its well-established antioxidant
potential.
2. MATERIALS AND METHODS
2.1 Preparation of the extract
Dehydrated basidiomes of Agaricus blazei Murill
(ABM) were bought from Ibema, Brazil. The Fine
powder was obtained by milling the basidiomes.
The powder was then subjected to extraction by
adding distilled water (100 ml) to 10 g each of
powdered basidiome and was kept under
agitation for 3 hours at a temperature of 28
°
C.
The remaining solids were removed through
filtration using Whatman filter paper (Size-1).
This process of extraction was repeated three
times whereas the remnants were lyophilized
and stored at -20°C.
2.2 Drugs and Chemicals
Rotenone (R8875 Sigma) and the other
chemicals which were used in this study were
procured from Sigma-Aldrich.
2.3 Phytochemical Screening
Freshly prepared Agaricus blazei Murill (ABM)
extract was put to different qualitative tests as
per procedures defined elsewhere [14] to
evaluate the existence of phytochemical
constituents such as carbohydrates, protein,
saponins, flavonoids, alkaloids, cardiac
glycosides, and anthraquinones, through virtue of
colour changes.
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
123
2.4 Selection of doses
The selection of doses of different drugs was
based on literature i.e., 273 mg/kg and 819
mg/kg for Agaricus blazei Murill [15] whereas 2.5
mg/kg for rotenone were used [16].
2.5 Animals
In this study, male wistar rats bearing the weight
of 200-220 g, were bought from the animal house
of University of Karachi. However, all the animals
were placed in the environment having optimum
temperature of 25-30 ˚C under 12\12-hour light-
dark cycle. Water ad libitum and free excess of
standard diet were provided to the animals.
2.5.1 Grouping of animals
Animals used in this study were divided in four
groups (n =6) as follows.
I: This group received sunflower oil 4 ml/kg, i.p.
II: This group received rotenone 2.5 mg/kg, i.p.
dissolved in sunflower oil 1 mg/2 ml
III: This group received Agaricus blazei Murill
273 mg/kg, p.o. and rotenone 2.5 mg/kg, i.p.
IV: Agaricus blazei Murill 819 mg/kg, p.o. and
rotenone 2.5 mg/kg, i.p.
All the groups of the animals received the doses
of above agents on daily basis for a period of 28
days.
2.6 Acute Toxicity of the Extract
Toxicity was assessed as per OECD-423
guidelines. Briefly, female Wistar rats weighing
between 200 to 220 g fasted overnight and a
single dose of 2000 mg/kg of the extract was
administered. For the detection of any signs of
toxicity the animals were observed for the next
24 hours. However, animals were carefully
monitored during the first 4 hours after the
administration of the extract [17].
2.7 Behavioral Assessment
Behavioral analyses were performed on 7
th
, 14
th
and 28
th
day of the study. All groups of the rats
were subjected to the following behavioral tests:
2.7.1 Catalepsy test
Catalepsy is a behavioral state in rodents in
which the animals become unable to correct the
externally imposed postures. In this study,
rotenone (2.5 mg/kg, i.p.) was used to induce the
catalepsy in rats. The period of catalepsy was
calculated in seconds. Catalepsy was measured
using a standard bar test. The bar was 9 cm
above the base. The front paws of the rat were
placed onto the 9 cm wooden bar and the
duration of retaining the forepaws on the
elevated bar until they touch the floor was noted
as the cataleptic score. The cutoff time of 180 s
was applied. The test was performed on 7
th
, 14
th
and 28
th
day of the study [18].
2.7.2 Exploratory behavior (rearing)
Rodents show exploratory behavior, including
rearing, whenever they are placed in a new
container. During rearing behaviour, the
forelimbs will touch the container wall. In this
study, small plexiglass cages size (30× 20× 30
cm) were used individually for each animal. The
rats were given 5 min habituation period before
the commencement of the test. After completion
of the habituation period, the number of rearing
was noted for the next 5 min. [19].
2.7.3 Rotarod motor coordination test
Rotarod was used to check the grip strength and
muscle rigidity of all animals. Rotarod test is
commonly used model for the assessment of
muscle coordination and motor function. Before
starting the therapy, to adjust rats on the rotarod
apparatus, each rat was trained. During the test,
each rat was placed on a rotating rod at a speed
of 25 rpm. A cutoff time of 180 seconds was
maintained during the experiment. Fall of time for
each animal was noted in seconds. The test was
performed on 7
th
, 14
th
and 28
th
day of the study
[18].
2.7.4 Locomotor activity
In this test, locomotor activity of animals was
evaluated on 7
th
, 14
th
and 28
th
day of the study by
using digital actophotometer having infrared
photocells for a period of 5 min. The values for
locomotor activity of animals were noted as
counts per 5 min [20].
2.8 Biochemical Estimation
2.8.1 Decapitation and homogenization
After the completion of behavioral tests, the
animals were anaesthetized and sacrificed by
decapitation for biochemical estimation.
Immediately after decapitation, the brains of the
animals were extract out, washed carefully with
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124
ice-cold normal saline, and homogenized in Tris
HCL. The supernatant fluid formed due to
homogenization was centrifuged for the period of
10 min at 10,000 ×g. The neuroprotective activity
of ABM extract was measured by estimating the
concentrations of brain antioxidant enzymes [12].
2.8.2 Lipid peroxidation assay (TBARS)
By measuring the byproduct of lipid peroxide i.e.
thiobarbituric acid reactive substances (TBARS),
lipid peroxide level can easily be measured. For
this purpose, aqueous solution of thiobarbituric
acid, sodium dodecyl sulphate and acetate buffer
(pH 3.5) were blended with (10 % w/v) tissue
homogenate. Red pigment was obtained after
heading the mixture at 95
C for 1 hour, which
was then extracted with n-butanolpyridine
mixture. The absorbance for this mixture was
noted at 532nm. Lipid peroxide level was
expressed as nmol malondialdehyde, whereas
tetramethoxypropane was used as an external
standard [21].
2.8.3 Reduced glutathione assay (GSH)
An assay of reduced glutathione was done by
precipitating 1 ml of both 10% trichloroacetic acid
(TCA) and the tissue homogenate. In addition,
5,5-dithio-bis-(2-nitrobenzoicacid)(DTNB)reagent
0.5 ml and 4 ml of phosphate solution were
added to the homogenized supernatant liquid.
The absorbance was noted at 412 nm [21].
2.8.4 Estimation of nitrite
Nitrite can be detected spectrophotometrically by
using Griess reagent. A mixture of Griess
reagent and brain homogenate was formed by
mixing them in equal proportions and incubated.
The absorbance of mixture was noted at 546 nm
[22].
2.8.5 Determination of protein
For the determination of protein, Lowry method
was used. Whereas bovine serum albumin was
used for standard curve determination [23].
2.9 Statistical Analysis
Data were expressed as ± standard error of
mean (SEM). One-way ANOVA followed by
Tukey’s post hoc test was applied for the
analysis of data.
3. RESULTS
3.1 Phytochemical Screening
The aqueous extract of ABM showed the
presence of pharmacologically important
phytoconstituents, which are summarized in
Table 1.
3.2 The Effect of ABM Extract on
Rotenone-induced Catalepsy in Rats
The result of one-way ANOVA showed extremely
significant difference between the four groups of
rats in duration of catalepsy (F
3, 8
=15.16, P =
0.001). The means of treatment groups viz.
Group III (ABME 273 mg/kg) and Group IV
(ABME 819 mg/kg) were compared with the
means of Group II (rotenone 2.5 mg/kg) using
Tukey’s post hoc test. However, it has been
observed that the activity of animals was
significantly reduced in the treatment groups
when compared with the rotenone group (Fig. 1).
3.3 The Effect of ABM Extract on
Exploratory Behavior (rearing) in Rats
In this behavioral test, there were significant
differences among four groups of the rats as
indicated by one-way ANOVA (F
3, 8
=6.15, P =
0.014). The means of the treatment groups viz.
Group III (ABME 273 mg/kg) and Group IV
(ABME 819 mg/kg) were compared with the
means of Group II (rotenone 2.5 mg/kg) using
Tukey’s post hoc test. However, it has been
observed that the activity of animals was
significantly increased in the treatment groups
when compared with the rotenone group (Fig. 2).
3.4 The Effect of ABM Extract on
Rotarod
Motor
Coordination
Test in
Rats
It this test, there were extremely significant
difference among the four groups of the rats as
indicated by one-way ANOVA (F
3, 8
=43.60, P =
0.001). The means of the treatment groups viz.
Group III (ABME 273 mg/kg) and Group IV
(ABME 819 mg/kg) were compared with the
means of Group II (rotenone 2.5 mg/kg) using
Tukey’s post hoc test. It has been observed the
fall off time in rotarod test was significantly
increased in the treatment groups when
compared with the rotenone group (Fig. 3).
3.5 The Effect of ABM Extract on
Rotenone-induced locomotor
Activity in Rats
The results of one-way ANOVA (F
3, 8
=53.92, P =
0.001) indicated a significant difference among
four groups of rats in actophotometer test. The
means of treatment groups viz. Group III (ABME
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
125
273 mg/kg) and Group IV (ABME 819 mg/kg)
were compared with the means of Group II
(rotenone 2.5 mg/kg) using Tukey’s post hoc
test. The results showed that the locomotor
activity was significantly increased in treatment
groups when compared with rotenone group (Fig.
4).
3.6 The Effect of ABM Extract on Brain
Nitrite, TBARS, GSH and Total
Proteins in Rats
In biochemical assay, a one-way ANOVA
indicated a significant difference among four
groups of rats in brain levels of nitrite (F
3, 20
=
39.59, P = 0.0001), TBARS (F
3, 20
= 14.17, P =
0.0001), GSH (F
3, 20
= 77.21, P = 0.0001) and
total proteins (F
3, 20
= 57.57, P = 0.0001). The
means of treatment groups viz. Group III (ABME
273 mg/kg) and Group IV (ABME 819 mg/kg)
were compared with the means of Group II
(rotenone 2.5 mg/kg) using Tukey’s post hoc
test. The results of the analysis reveal that the
levels of nitrite and TBARS were significantly
decreased while the levels of reduced GSH and
total proteins were significantly increased in
treatment groups when compared with rotenone
group (Figs. 5-8).
Table 1. Phytochemical analysis of the aqueous extract of Agaricus blazei Murill
Phytochemical
Test
Observation
Carbohydrates
Barfoed’s and Molisch’s test +
+
Protein Biuret test +
Flavonoids Pew’s test
Shibita’s reaction test
+
+
Saponins Frothing test +
Alkaloids Mayer’s and Dragendorff’s test +
+
Anthraquinones Borntrager’s test ˗˗
Cardiac glycosides General test ˗˗
(+ present; -- absent)
Fig. 1. Effect of ABM extract on rotenone-induced catalepsy in rats.
The values are mean ± SEM
a
p < 0.05,
b
p < 0.01,
c
p < 0.001 when compared with the control group,
d
p < 0.05,
e
p < 0.01,
f
p < 0.001 when
compared with the rotenone group;
(ANOVA followed by Tukey’s post hoc test)
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
126
Fig. 2. Effect of ABM extract on exploratory behavior (rearing) in rats
The values are mean ± SEM;
a
p < 0.05,
b
p < 0.01,
c
p < 0.001 when compared with the control group,
d
p < 0.05,
e
p < 0.01,
f
p < 0.001 when compared with the rotenone group; (ANOVA followed by Tukey’s post hoc test)
Fig. 3. Effect of ABM extract on rotarod motor coordination test in rats
The values are mean ± SEM.;
a
p < 0.05,
b
p < 0.01,
c
p < 0.001 when compared with the control group.,
d
p < 0.05,
e
p < 0.01,
f
p < 0.001 when compared with the rotenone group; (ANOVA followed by Tukey’s post hoc test).
Fig. 4. Effect of ABM extract on locomotor activity in rats
The values are mean ± SEM;
a
p < 0.05,
b
p < 0.01,
c
p < 0.001 when compared with the control group,
d
p < 0.05,
e
p < 0.01,
f
p < 0.001 when compared with the rotenone group; (ANOVA followed by Tukey’s post hoc test)
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
127
Fig. 5. Effect of ABM extract on nitrite levels in the rat brain
The values are mean ± SEM
a
p<0.05,
b
p<0.01,
c
p<0.001 when compared with the control group,
d
p<0.05,
e
p<0.01,
f
p<0.001 when compared
with the rotenone group; (ANOVA followed by Tukey’s post hoc test)
Fig. 6. Effect of ABM extract on lipid peroxidation in the rat brain
The values are mean ± SEM;
a
p<0.05,
b
p<0.01,
c
p<0.001 when compared with the control group,
d
p<0.05,
e
p<0.01,
f
p<0.001 when compared with the rotenone group; (ANOVA followed by Tukey’s post hoc test)
Fig. 7. Effect of ABM extract on reduced glutathione levels in the rat brain.
The values are mean ± SEM;
a
p<0.05,
b
p<0.01,
c
p<0.001 when compared with the control group,
d
p<0.05,
e
p<0.01,
f
p<0.001 when compared with the rotenone group; (ANOVA followed by Tukey’s post hoc test)
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
128
Fig. 8. Effect of ABM extract on total protein levels in the rat brain.
The values are mean ± SEM;
a
p<0.05,
b
p<0.01,
c
p<0.001 when compared with the control group,
d
p<0.05,
e
p<0.01,
f
p<0.001 when compared with the rotenone group; (ANOVA followed by Tukey’s post hoc test)
4. DISCUSSION
Parkinson’s Disease, a chronic
neurodegenerative disorder which may be due to
the reduction in dopaminergic neurons present in
the region of substantia nigra pars compacta.
Whereas, several pathologies such as
mitochondrial dysfunction, oxidative stress,
protein accumulation like a-synuclein and
apoptosis are involved in this disease. However,
the most important pathology of PD has been
oxidative stress [24].
Rotenone-induced catalepsy, exploratory
behavior, rotarod test and locomotor activity are
most frequently used models for animal to
evaluate neurodegenerative disorders. A specific
complex I inhibitor i.e., rotenone which is a
common herbicide, reproduces Parkinsonian
signs and symptoms in rodents [18]. Many
research studies have exhibited that systemic
administration of rotenone can lead to the
degradation of dopaminergic neurons in the
nigrostriatal pathway that progresses the
development of behavioural, neurochemical and
pathological events of PD [25].
The results of rotenone-induced catalepsy
showed that Agaricus blazei Murill provides
significant ameliorative effect on Parkinson’s
disease in rats. The effect of Agaricus blazei
Murill extract on rearing, muscle rigidity and
locomotor activity of rats was also evaluated
because earlier studies show that the patient
suffering from PD reflects loss of brain motor
coordination and becomes unable to maintain
normal limb posture [26]. Efficient locomotor
activity was observed among the animals treated
with Agaricus blazei Murill extract at the doses of
273 and 819 mg/kg when compared to control
group which provided more evidence of the
ameliorative effect of Agaricus blazei Murill on
PD.
Dysfunction of mitochondrial complex-1
generates oxidative stress and plays an
imperative role in the pathogenesis of
Parkinson’s Disease [24]. Agaricus blazei Murill
is a well-recognized antioxidant mushroom. [27].
Considering the potent antioxidant profile of
Agaricus blazei, it is assumed that the beneficial
effect of Agaricus blazei Murill in aforesaid
models of PD could be due to its lessening effect
on the oxidative stress in the brain. To confirm
the assumption, the levels of reduced
glutathione, total protein, nitric oxide and
thiobarbituric acid reactive substances (TBARS)
were estimated in the animal brain.
Glutathione peroxidase is one of the important
neuroprotective enzymes in the brain. It acts as a
scavenger of H
2
O
2
produced by cellular
metabolism besides balancing the composition
and disintegration of H
2
O
2
in normal conditions.
The decreased level of glutathione is the limiting
factor in the elimination of H
2
O
2
. However, in
Parkinson’s disease, glutathione is reduced
extensively in the substantia nigra because of
neuronal loss [28]. It has been reported that
excess stimulation of neurons by the glutamic
acid and significant activation of macrophages
through nitric oxide can promote the toxicity of
neuronal cells in the brain [29]. The production of
nitric oxide in biological materials can be
assessed by the determination of nitrite. In this
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129
study, the level of nitric oxide in the brain was
detected spectrophotometrically by using Griess
reagent [30]. The biomarker of oxidative stress is
lipid peroxidation which was determined by
measuring the byproduct of lipid peroxide i.e.,
thiobarbituric acid reactive substances (TBARS).
Lipid peroxidation ensues because of the attack
on double bonds of arachidonic acid and
unsaturated fatty acid. This generates the
radicles of lipid peroxyl, which initiate a series of
additional strikes on other unsaturated fatty acids
that can leads to the oxidative degradation of
polyunsaturated fatty acids and its incidence in
bio membranes results in impaired structural
integrity, inactivation of several membrane-bound
enzymes, impaired membrane function and
reduced fluidity. Earlier studies show that in the
substantia nigra of PD patients the levels of lipid
peroxidation product is significantly increased
[31]. In the current study, analogous results have
been found in the brain homogenate of rotenone
treated rats. Rotenone animal group exhibited a
steady decline in GSH and total protein levels
and a substantial rise in the levels of nitrite and
TBARS in the animal brain when compared to
the control group. Two different doses of
aqueous extract of Agaricus blazei Murill (273
and 819 mg/kg, p.o.) were used in the rotenone-
induced PD models. Both doses were found to
be significant in decreasing oxidative stress in
the animal brain by increasing the brain levels of
reduced GSH and total proteins and decreasing
the levels of nitrite and TBARS.
5. CONCLUSION
The results of rotenone-induced catalepsy,
exploratory behavior, rotarod test and locomotor
activity showed that Agaricus blazei Murill exerts
a significant ameliorative effect on Parkinson’s
disease in rats. Biochemically, both doses were
found to be significant in decreasing oxidative
stress in the animal brain by increasing the brain
levels of reduced GSH and total proteins and
decreasing the levels of nitrite and TBARS. Thus,
we conclude that Agaricus blazei Murill
possesses antiparkinsonian activity in rats. This
activity of the mushroom could be due to its
antioxidant potential. The mushroom can be
used alone or in combination with other
antiparkinsonian drugs. We believe that
consumption of the mushroom will be highly
fruitful for the PD patients.
CONSENT
It is not applicable.
ETHICAL APPROVAL
This study was reviewed and approved by
institutional ethical committee of Ziauddin
University, Karachi.
COMPETING INTERESTS
Authors have declared that no competing
interests exist.
REFERENCES
1. Hauser DN, Hastings TG. Mitochondrial
dysfunction and oxidative stress in
Parkinson's disease and monogenic
parkinsonism. Neurobiol Dis. 2013;51:35-
42.
2. Wang H, Fu Z, Han C. The medicinal
values of culinary-medicinal royal sun
mushroom (Agaricus blazei Murrill). Evid
Based Complement Alternat Med; 2013.
3. Ramoutsaki IA, Ramoutsakis IA,
Papadakis CE, Helidonis ES. Therapeutic
methods used for otolaryngological
problems during the Byzantine period. Ann
Otol Rhinol Laryngol. 2002;111(6):553-7.
4. Akiyama H, Endo M, Matsui T, Katsuda I,
Emi N, Kawamoto Y, Koike T, Beppu H.
Agaritine from Agaricus blazei Murrill
induces apoptosis in the leukemic cell line
U937. Biochim Biophys Acta.
2011;1810(5):519-25.
5. Giavasis I. Bioactive fungal
polysaccharides as potential functional
ingredients in food and nutraceuticals. Curr
Opin Biotechnol. 2014;26:162-73.
6. Smiderle FR, Alquini G, Tadra-Sfeir MZ,
Iacomini M, Wichers HJ, Van Griensven
LJ. Agaricus bisporus and Agaricus
brasiliensis (1→ 6)-β-d-glucans show
immunostimulatory activity on human THP-
1 derived macrophages. Carbohydr Polym.
2013;94(1):91-9.
7. Yamamoto KA, Galhardi LC, Rincão VP,
de Aguiar Soares S, Vieira ÍG, Ricardo
NM, Nozawa C, Linhares RE. Antiherpetic
activity of an Agaricus brasiliensis
polysaccharide, its sulfated derivative and
fractions. Int J Biol Macromol. 2013;52:9-
13.
8. Alves MJ, Ferreira IC, Froufe HJ, Abreu
RM, Martins A, Pintado M. Antimicrobial
activity of phenolic compounds identified in
wild mushrooms, SAR analysis and
docking studies. J Appl Microbiol.
2013;115(2):346-57.
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
130
9. Valverde ME, Hernández-Pérez T,
Paredes-López O. Edible mushrooms:
improving human health and promoting
quality life. Int J Microbiol. 2015.
10. de Miranda AM, Ribeiro GM, Cunha AC,
Silva LS, dos Santos RC, Pedrosa ML,
Silva ME. Hypolipidemic effect of the
edible mushroom Agaricus blazei in rats
subjected to a hypercholesterolemic diet. J
Physiol Biochem. 2014;70(1):215-24.
11. Soares AA, de Oliveira AL, Sá-Nakanishi
AB, Comar JF, Rampazzo AP, Vicentini
FA, Natali MR, Gomes da Costa SM,
Bracht A, Peralta RM. Effects of an
Agaricus blazei aqueous extract
pretreatment on paracetamol-induced
brain and liver injury in rats. Biomed Res
Int. 2013.
12. Koppula S, Kumar H, More SV, Kim BW,
Kim IS, Choi DK. Recent advances on the
neuroprotective potential of antioxidants in
experimental models of Parkinson’s
disease. Int J Mol Sci. 2012;13(8):10608-
29.
13. Hakime-Silva RA, Vellosa JC, Khalil NM,
Khalil OA, Brunetti IL, Oliveira OM.
Chemical, enzymatic and cellular
antioxidant activity studies of Agaricus
blazei Murrill. An Acad Bras Cienc.
2013;85(3):1073-82.
14. Aziz MA. Qualitative phytochemical
screening and evaluation of anti-
inflammatory, analgesic and antipyretic
activities of Microcos paniculata barks and
fruits. J Integr Med. 2015;13(3):173-84.
15. Ni WY, Wu MF, Liao NC, Yeh MY, Lu HF,
Hsueh SC, Liu JY, Huang YP, Chang CH,
Chung JG. Extract of medicinal mushroom
Agaricus blazei Murill enhances the non-
specific and adaptive immune activities in
BALB/c mice. In vivo. 2013;27(6):779-86.
16. Javed H, Azimullah S, Khair SB, Ojha S,
Haque ME. Neuroprotective effect of
nerolidol against neuroinflammation and
oxidative stress induced by rotenone. BMC
Neurosci. 2016;17(1):1-2.
17. Jonsson M, Jestoi M, Nathanail AV,
Kokkonen UM, Anttila M, Koivisto P,
Karhunen P, Peltonen K. Application of
OECD Guideline 423 in assessing the
acute oral toxicity of moniliformin. Food
Chem Toxicol. 2013;53:27-32.
18. Chen Y, Zhang DQ, Liao Z, Wang B, Gong
S, Wang C, Zhang MZ, Wang GH, Cai H,
Liao FF, Xu JP. Anti-oxidant polydatin
(piceid) protects against substantia nigral
motor degeneration in multiple rodent
models of Parkinson’s disease. Mol
Neurodegener. 2015;10(1):1-4.
19. Cannon JR, Tapias V, Na HM, Honick AS,
Drolet RE, Greenamyre JT. A highly
reproducible rotenone model of
Parkinson's disease Neurobiol Dis.
2009;34(2):279-90
20. Sharma N, Nehru B. Beneficial effect of
vitamin E in rotenone induced model of
PD: behavioural, neurochemical and
biochemical study. Exp Neurobiol.
2013;22(3):214.
21. Aslam M, Sial AA. Neuroprotective effect
of ethanol extract of leaves of Malva
parviflora against amyloid-β-(Aβ-)
mediated Alzheimer’s disease. Int Sch Res
Notices; 2014.
22. Radenović L, Selaković V, Janać B,
Todorović D. Effect of glutamate
antagonists on nitric oxide production in rat
brain following intrahippocampal injection.
Arch Biol Sci. 2007;59(1):29-36.
23. Ledoux M, Lamy F. Determination of
proteins and sulfobetaine with the folin-
phenol reagent. Ana Biochem.
1986;157(1):28-31.
24. Friedlich AL, Smith MA, Zhu X, Takeda A,
Nunomura A, Moreira PI, Perry G.
Oxidative stress in Parkinson’s disease.
Open Pathol J. 2009;3(1):38-42
25. Alves MJ, Ferreira IC, Froufe HJ, Abreu
RM, Martins A, Pintado M. Antimicrobial
activity of phenolic compounds identified in
wild mushrooms, SAR analysis and
docking studies. J Appl Microbiol.
2013;115(2):346-57.
26. Bais S, Gill NS, Kumar N. Neuroprotective
effect of Juniperus communis on
chlorpromazine induced Parkinson disease
in animal model. Chinese J Biol; 2015.
27. Niranjan R. The role of inflammatory and
oxidative stress mechanisms in the
pathogenesis of Parkinson’s disease:
focus on astrocytes. Mol Neurobiol.
2014;49(1):28-38.
28. García-Robledo E, Corzo A, Papaspyrou
S. A fast and direct spectrophotometric
method for the sequential determination of
nitrate and nitrite at low concentrations in
small volumes. Marine Chem.
2014;162:30-6.
29. Lopes MÂ, Meisel A, Carvalho FD, de
Lourdes Bastos M. Neuronal nitric oxide
synthase is a key factor in doxorubicin-
induced toxicity to rat-isolated cortical
neurons. Marine Chem. 2011;19(1):14-22.
Aslam et al.; JPRI, 33(33B): 121-131, 2021; Article no.JPRI.69598
131
30. Ricart-Jane D, Llobera M, Lopez-Tejero
MD. Anticoagulants and other preanalytical
factors interfere in plasma nitrate/nitrite
quantification by the Griess method. Nitric
Oxide. 2002; 6(2):178-85.
31. Bhangale JO, Acharya SR. Anti-Parkinson
activity of petroleum ether extract of Ficus
religiosa (L.) leaves. Adv Pharmacol Sci;
2016.
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... 20−23 Medicinal mushrooms (basidiocarps/mycelia extracts or isolated bioactive compounds) have gained attention due to their protective effects against neurodegenerative diseases. 24,25 Ganoderma lucidum extract protected the death of DA neurons in a dose-dependent manner in the PD model and downregulated the expression of proinflammatory cytokines like tumor necrosis factor-alpha (TNF-α) and interleckin-1β (IL-1β). 26 Sanghuangprous vaninii (S. vaninii), also called "sanghuang" in China, is a well-known mushroom within the Hymenochaetaetaceae family. ...
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