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Journal of Drug Research and Development
ISSN 2470-1009 | Open Access
J Drug Res Dev | JDRD
1
RESEARCH ARTICLE
Hericium erinaceus Mycelium Exerts Neuroprotecve Eect in Parkinson’s
Disease-in vitro and in vivo Models
Pao-Pao Yang1,2, Chih-Yung Lin2, Tzu-Yin Lin2, and Win-Chin Chiang2,*
1Instute of Biotechnology and Pharmaceucal Research, Naonal Health Research Instutes, Miaoli County, Taiwan, Republic of China
2JOWIN BIOPHARMA Inc, Taiwan, Republic of China
Received: 20 Feb, 2020 | Accepted: 13 Mar, 2020 | Published: 20 Mar, 2020
Volume 6 - Issue 1
*Corresponding author: Win-Chin Chiang, JOWIN BIOPHARMA Inc, Xizhi, New Taipei City, Taiwan, 9F-12, No. 97, Sec. 1, Xintai 5th Road, Xizhi
Dist, New Taipei City, Taiwan (22175), E-mail: winchiang@jowinbio.com
Citaon: Yang PP, Lin CY, Lin TY, Chiang WC (2020) Hericium erinaceus Mycelium Exerts Neuroprotecve Eect in Parkinson’s Disease-in vitro and in
vivo Models. J Drug Res Dev 6(1): dx.doi.org/10.16966/2470-1009.150
Copyright: © 2020 Yang PP, et al. This is an open-access arcle distributed under the terms of the Creave Commons Aribuon License,
which permits unrestricted use, distribuon, and reproducon in any medium, provided the original author and source are credited.
Abstract
Hericium erinaceus (H.E.) is a well-known edible and folk medicinal fungi in Japan, China and other Asian countries without harmful eects. It has
been recognized that this unique mushroom is capable of keeping the brain healthy, supporng the immune system to help prevent gastric cancer
and other diseases, boosng mood and concentraon, decreasing inammatory processes in the body. But more scienc researchers are needed
to conrm its nutrional and medicinal eects. In the present study, we invesgated the eects of Hericium erinaceus mycelium (H.E. mycelium)
against 1-methyl-4-phenylpyridinium(MPP+)-induced neurotoxicity in PC12 cells and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced
Parkinsonian mice. In the cell viability results, treatment with H.E.mycelium increased the cell viability in MPP+-treated cells and induced anoxidant
acvity in PC12 cells. H.E. mycelium also reduced MPTP-induced loss of dopamine concentraon level and tyrosine hydroxylase (TH) posive cells in
mice. Our results suggest that H.E.mycelium performs signicant protecon of dopaminergic neuron under severe condions and is very eecve in
the treatment of damaged neuron in the brain to recover in the case of Parkinson’s disease.
Praccal applicaons: In this arcle, we provide science-based evidence related to H.E.mycelium to be a potenal eecve material for the
treatment and prevenon of Parkinson’s disease.
Keywords: Hericium erinaceus mycelium; Dopamine; Neuroprotecve; Parkinson’s disease; Tyrosine hydroxylase
Introduction
Parkinson’s disease (PD) is one of the most common progressive
neurodegenerative disease that is characterized by the loss of
dopaminergic neurons in the Substantia Nigra par compacta (SNpc)
region of the brain [1], which results in motor problems including
bradykinesia, akinesia, muscular rigidity, resting tremor, and postural
instability [2]. In the disease model of PD, the involvement of the
drug 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP),
is most widely used among animal models of Parkinson’s disease
[3]. e MPTP is metabolized into the toxic cation 1-methyl-4-
phenylpyridinium ion (MPP+) by the enzyme monoamine oxidase
B, which causes the neurotoxic eect to impair the dopaminergic
nigrostriatal neurons [4].
Hericium erinaceus (H.E.) a well reputed edible mushroom, also
known as monkey head mushroom in Chinese, Yamabushitake
in Japanese or Lion’s mane mushroom in English, has been widely
reported to use as food and folk medicine in Japan, [5] China and
other Asian countries without harmful eects [6]. Several evidences
demonstrated that it possesses a wide range of benets, such as
anticancer [7,8], antimicrobial [9], antioxidant [10], anti aging
[11], anti-hyperglycemic [12], anti-hyperlipidemic activity [13],
gastroprotective [14], immunomodulating and neuroprotective
activity (Alzheimer’s Disease and Parkinson’s Disease) [15-17],
protection of neuropathic pain [18], depressive symptoms [19] and
presbycusis [20].
As the mycelium is inoculated in grain spawn, lion’s mane
mushroom grows in large snowball-like formations, which is called
the fruiting body. Hericenones, the benzyl alcohol derivatives
with simple fatty acids, only exist in the fruiting body. A group of
erinacines (erinacines A-K and P-S) which are diterpenoid derivatives
have been identied only from the mycelium, and erinacine A is
most rich in the mycelium [21]. It was demonstrated that eight of
erinacines (A-I) could enhance nerve growth factor (NGF) release
[21] and the erinacenes are more potent inducers of NGF synthesis
than hericenones. ey not only have an enhancing eect on NGF
synthesis in astroglial cells in vitro but also can increase both NGF and
catecholamine content in the hippocampus of rats [11]. e increased
amount of NGF, in turn, enhances neuronal survival in dierent brain
regions and substantially improves animal behavioral activity.
Sci Forschen
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Citaon: Yang PP, Lin CY, Lin TY, Chiang WC (2020) Hericium erinaceus Mycelium Exerts Neuroprotecve Eect in Parkinson’s Disease-in vitro
and in vivo Models. J Drug Res Dev 6(1): dx.doi.org/10.16966/2470-1009.150 2
Journal of Drug Research and Development
Open Access Journal
mycelium is the concentration of H.E. mycelium required to inhibit
50% of the DPPH free radical. Likewise, the IC50 value of Ganoderma
lucidum was so dened. e percent inhibition of DPPH free radical
by a test sample calculated by using the following formula:
% of inhibition = [(dc-dt)/dc] × 100
Where dc was the absorbance of control reaction and dt was the
absorbance in presence of test or standard sample.
Animals and treatment
Adult male C57BL/6 Narl mice weighing 20-30 g (8-12 weeks old)
were purchased from the National Laboratory Animal Center (Taipei,
Taiwan) and used in this study. Mice were kept at constant room
temperature (20-22°C) and humidity (50%-70%) with a 12 h light/
dark cycle (7:00-19:00) in the Animal Centers of the National Yang-
Ming University, Taiwan. Standard diet and water were available Ad
libitum during the experiment. e care of animals was carried out in
accordance with institutional and international standards (Principles
of Laboratory Animal Care, NIH); and all experiments were approved
by the Institutional Animal Care and Use Committees of the National
Yang-Ming University, Taiwan (IACUC No. 1050306). All studies
involving animals were conducted in accordance with the ARRIVE
guidelines [22,23].
Five groups (een mice in each group) were randomly assigned
to a control group (saline, i.p.) plus H2O (p.o.), MPTP group (20 mg/
kg, i.p; Tokyo chemical Industry, TCI) plus H2O (p.o.) and MPTP (20
mg/kg, i.p.) plus H.E. mycelium groups (0.1 g/kg, 0.3 g/kg and 1 g/kg.,
p.o.). Mice were received intraperitoneal injection of saline or MPTP
once a day in the beginning 5 consecutive days and oral administration
of H2O or H.E. mycelium in the duration of 30-day period. e control
group of animal received an equivalent volume of saline. e animals
were sacriced 30 days aer the last treatment of ve groups and
then brains were dissected to the le and right cerebrum. e right
cerebrum was removed to determine immunohistochemistry for
tyrosine hydroxylase and the le cerebrum of striatum was rapidly
removed to determine concentration of dopamine neurotransmitter.
Determination of the concentration of dopamine
neurotransmitter
e mice were sacriced under pentobarbital aer completion of
the treatment. e le cerebrum of striatum was quickly removed and
homogenized in a stock solution containing 0.1 M HClO4, 0.1 mM
EDTA, 0.1 mM Na2S2O5 and centrifuged at 13,000 rpm for 10 min
at 4°C. e supernatant was harvested, ltered with 0.45 µm pore-
size lters, and reserved at -80°C until analysis. e concentration
of dopamine was determined by using High-Performance Liquid
Chromatography (HPLC) with electrochemical detection.
High-performance liquid chromatography assay for
neurotransmitter
HPLC electrochemical detection was performed to quantify the
concentration of dopamine. e HPLC system consisted of a pump
(BAS PM-92E; Bioanalytical Systems, West Lafayette, IN, USA),
a refrigerated microsampler (CMA/200) and a sample injector
(CMA/240) with a 20 µL loop (CMA, Stockholm, Sweden), and a
digital amperometric electrochemical detector (Decade II; Antec
Leyden BV, Zoeterwoude, e Netherlands). For the determination
of the dopamine concentration, 15 µL samples were injected into
the HPLC system. e mobile phase consisted of 0.74 mM sodium-
1-Octanesulfaoate (SOS), 100 mM phosphate sodium salt, 0.027 mM
EDTA, 2 mMKCl, 125 mL methanol, which was delivered at a ow
e aim of this present study was to explore the neuro protective
eects of Hericium erinaceus mycelium (H.E. mycelium) using
MPP+-treated PC12 cells or MPTP-induced PD mouse model that
is associated with protection against loss of the neurotransmitter or
dopaminergic neuron in vitro and in v ivo.
Materials and Methods
Preparation of Hericium erinaceus mycelium (H.E.
mycelium)
H.E. mycelium powder (mesh size#100, RH6408) were obtained
from FUNGUS BIOTECH, Co. Ltd. Yilan, Taiwan, where toxin-free
and pesticide-free of Hericium erinaceus solid state fermentation was
exercised and the mycelium was collected aerward and dried to
the moisture content of less than 7%. e H.E. mycelium yellowish
powder was then further grinded into smaller particles through a
spiral jet mill (OM2 micronizer, Sturtevant Inc. Hanover, MA
USA) to induce the cell wall-broken effect with a particle size
distribution of D75<50 µm at FORMOSAN NANO BIOLOGY
Co. Ltd, Taichung, Taiwan. e cell wall-breaking technology greatly
contributed to the increased release rate of active ingredients from the
ne H.E. Mycelium particle powder.
Cell viability
e MTS assay is a colorimetric method and usually used to assess
cell proliferation, cell viability and cytotoxicity. Its protocol is based
on the reduction of the MTS tetrazolium compound by viable cells to
generate a colored formazan dye that is soluble in cell culture media.
PC12 cell is a cell line derived from a phenochromocytoma of the
rat adrenal medulla and was used in this study. e PC12cells were
maintained at 3*104cells /well in 96 wells plate with 100 µL of DMEM
at 37°C in an incubator containing 5% carbon dioxide for 24 h. Briey,
aer cells had attached, cells were treated with MPP+ for 72 h in the
presence of ethanol extracts of test samples which was in 0.4% DMSO
(Dimethyl Sulfoxide) solution. Aerward, MTS solution was added to
each cell well and made it into a colored solution. e whole process
was performed triplicate. e absorbance of the colored solution
in each cell was measured at 570 nm using a microplate reader. To
assess the neuroprotective eects of H.E. mycelium on the PC12 cells
with MPP+-induced toxicity, the cells were treated with dierent
concentration of H.E. mycelium at 8, 40, 200, 1000 µg/mL, respectively,
and dierent concentrations of Ganoderma lucidum (Reishi) at 8, 40,
200, 1000 µg/mL, respectively, with addition of 10 mM MPP+ solution
to reach the nal concentration of 1 mM MPP+ in the cell. Cell viability
was assessed 72 h later by measuring the absorbance of the colored
solution. e survival rate of the control group was normalized as the
basis for that of the other groups to compare and calculate.
DPPH scavenging assay
e free radical scavenging activity of the tested extract was
performed by using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) which
was at 0.1 mM Methanol solution. e extract fractions at dierent
concentrations were prepared with distilled water/ethanol (50:50).
e antioxidant standard compound, ascorbic acid, at 1 mg/mL was
used as positive standard for the comparison purpose. A fresh stock
solution of the standard compound was prepared before each analysis.
e absorbance changes in color from deep purple to light yellow at
517 nm were measured by using a spectrophotometer aer 30 min of
reaction. e solution of DPPH at 0.1 mM was used as a control.
To express the radical scavenging activity, the IC50 parameter was
employed and it is dened as the concentration of substrate that
brings about 50% loss of the DPPH free radical. e IC50 value of H.E.
Sci Forschen
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Citaon: Yang PP, Lin CY, Lin TY, Chiang WC (2020) Hericium erinaceus Mycelium Exerts Neuroprotecve Eect in Parkinson’s Disease-in vitro
and in vivo Models. J Drug Res Dev 6(1): dx.doi.org/10.16966/2470-1009.150 3
Journal of Drug Research and Development
Open Access Journal
rate of 500 µL/min. A reversed-phase C-18 column (100 × 4.6 mm,
2.6 µm) was used for sample separation. The applied potential of
the glassy carbon electrode was 650 mV to the reference electrode
(Ag/AgCl), and the range setting was 5 nA for the determination
of neurotransmitters. The data acquisition and analysis were
performed using EZChrom software (Scientific Software, San
Ramon, CA, USA).
Immunohistochemistry (IHC) for Tyrosine Hydroxylase
(TH) in mouse brain
For measuring tyrosine hydroxylase immunohistochemistry, the
mice were anesthetized with pentobarbital aer completion of the
treatment. e right cerebrum was removed and soaked at 4°C in the
paraformaldehyde (PFA) alone, and maintained in 30% sucrose at 4°C
until they sank. e cerebrum was sectioned at 10 µm. All sections
were stained for tyrosine hydroxylase measurements.
Data analysis and statistical assessment
Data were expressed as the mean ± SEM. Analysis of variance was
used to access the statistical signicance for repeated measures of the
data, and the dierences among the individual mean values in dierent
groups were analyzed by ANOVA followed by the Newman-Keuls test.
e dierences were considered to be signicant at p<0.05.
Results
DPPH scavenging activity (antioxidant activity)
e DPPH radical scavenging assay was determined
spectrophotometrically. e radical scavenging capacities of the
tested extract were performed by DPPH assay and results were shown
in gure 1. DPPH assay was one of the most widely and commonly
used methods for screening antioxidant activity of plant or mushroom
extracts. In the gure 1, DPPH scavenging activities of H.E. mycelium,
and reference antioxidants capacities of Ganoderma lucidum (Reishi)
obtained from FUNGUS BIOTECH, Co. Ltd., Yilan, Taiwan and
ascorbic acid were analyzed in this study. e concentrations of H.E.
mycelium at the range of 62.5 and 1000 µg/mL exhibited positive
DPPH scavenging activities in a concentration-dependent manner as
had shown in gure 1. e higher the concentration of H.E. mycelium
at, the more the DPPH radical scavenging activity. As compared,
Ganoderma lucidum showed a relatively lower radical scavenging
eect than H.E. mycelium. e radical scavenging eect of the H.E.
mycelium can also be observed by comparing IC50 value. A lower IC50
value indicates a higher antioxidant activity. e IC50 value of the H.E.
mycelium was 217.2 µg/mL, compared to that of Ganoderma lucidum with
550.4 µg/mL. It indicated H.E. mycelium had a better radical scavenging
eect than Ganoderma lucidum. Overall, these ndings support that H.E.
mycelium exerted a good antioxidant activity in PC12 cells.
H.E. mycelium protected MPP+- induced neurotoxicity in
PC12 cell
e protective ability of H.E. mycelium from the cytotoxicity of
MPP+ in PC12 cells was measured by using the MTS assay. e results
of the measurement, as shown in gure 2, revealed the decrease of cell
viability of PC12 cells aer exposure to 1 mM MPP+ for 72 h. In the
presence of 1 mM MPP+, H.E. mycelium at the concentration range
of 40, 200 and 1000 µg/mL showed a statistically signicant protective
Figure 1: DPPH (1-Diphenyl-2-picrylhydrazyl) scavenging eect of H.E.
mycelium and Ganoderma lucidum. Ascorbic acid (Vitamin C) was
used as posive control. Data are presented as means ± SEM.
Figure 2: H.E. mycelium protected MPP+ induced neurotoxicity in
PC12 cells. PC12 cells were treated with H.E. mycelium or Ganoderma
lucidum with MPP+. Cell viability was measured by MTS assay. Data
are presented as means ± SEM. One way ANOVA and Newman-Keuls
tests were used to analyze the data. *P<0.05; **p<0.01; ***p<0.001
compared to 0.4% DMSO with 1 mM MPP+.
Figure 3: H.E. mycelium prevented MPTP-induced level of dopamine
concentraon. H.E. mycelium (0.1, 0.3 and 1 g/kg) and MPTP on the
concentraon of dopamine level in striatum. Data are presented as
means ± SEM (n=10-15). One way ANOVA and Newman-Keuls tests
were used to analyze the data. **p<0.01; ***p<0.001 compared to
when compared with control; ##P<0.01 when compared to MPTP.
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Citaon: Yang PP, Lin CY, Lin TY, Chiang WC (2020) Hericium erinaceus Mycelium Exerts Neuroprotecve Eect in Parkinson’s Disease-in vitro
and in vivo Models. J Drug Res Dev 6(1): dx.doi.org/10.16966/2470-1009.150 4
Journal of Drug Research and Development
Open Access Journal
eect in a concentration dependent manner. A high concentration
exerted a better neuroprotective eect. But, Ganoderma lucidum
showed a signicant protective eect in the PC12 cells only at high
concentration of 1000 µg/mL, as shown in gure 2. It can be concluded
that H.E. mycelium exerted a very good protective eect against the
MPP+ neurotoxicity in PC 12 cells.
H.E. mycelium prevented MPTP-induced reduction of
dopaminergic neuron
e protective eect of H.E. mycelium against MPTP-induced toxic
damage on dopaminergic neuron can be interpreted by looking at the
reduction of decreased dopamine level in the striatum of the brain aer
H.E. mycelium treatment. e dopamine concentration was measured
by using HPLC. From gure 3, one can recognize that ve injections
of MPTP (20 mg/kg, i.p.) reduced to about 46% of dopamine level of
striatum in mice, compared to the control group. As expected, the
oral administration of H.E. mycelium at 0.3 and 1 g/kg increased the
dopamine level to 56% and 79%, respectively, which indicated that the
protective eect of H.E. mycelium was 20% and 70% at 0.3 and 1.0 g/
kg, respectively, as compared to the MPTP group [24]. ese results
demonstrated that H.E. mycelium exerted the protective eect against
the MPTP-induced dopamine neuron damage.
H.E. mycelium prevented MPTP-induced death of tyrosine
hydroxylase(TH)-positive neuron in striatum in mice
e enzyme tyrosine hydroxylase (TH) converts the amino acid
L-tyrosine into 3,4-dihydroxyphenylalanine (L-DOPA) which then
converts to dopamine by decarboxylation and then norepinephrine
and epinephrine are produced in series in the pathway in both central
and peripheral nervous systems [25]. e biosynthetic of dopamine
pathway is illustrated in gure 4. erefore, the activity of tyrosine
hydroxylase can inuence the dopamine level of striatum in brain
tissue and the reduction of tyrosine hydroxylase activity represents
the damage of dopaminergic neurons in the brain. Figure 5 shows a
large number of TH-positive cells in the control group and MPTP
caused 63% reduction in TH-positive cells in mice compared with
the control group. Aer treatment of H.E. mycelium at 0.1, 0.3 and
1 g/kg in the MPTP-induced mice, it improved the TH density in the
substantia nigra area of the brain by 18%, 78% and 100% at these three
concentrations respectively, compared with the MPTP damage group.
ese ndings conrmed that H.E. mycelium exerted protection from
MPTP-induced death of TH-positive neurons in corpus striatum of mice.
Discussion
In this present study we demonstrates that the neuroprotective
eect of H.E. mycelium against MPP+-induced toxicity in neuronal
PC12 cell lines and MPTP-induced striatal dopamine neuron damage
in mice model. Our data strongly suggests that H.E. mycelium intake
could be a potential treatment of PD (Parkinson’s disease).
Parkinson’s disease (PD) is characterized by a progressive
degeneration of dopaminergic neurons in the substantia nigra pars
compacta (SNpc) region of the brain [1], and N-methyl-4-phenyl-
1,2,3,6-tetrahydropyridine (MPTP) has been widely used to induce
PD in the animal model. MPTP induces PD through the death of
dopaminergic cell by its active metabolite, 1-methyl-4-phenyl-2,3,-
dihydropyridinium (MPP+) [3]. By using MPP+ in PC12 cells, as a
cellular model of PD, we have elucidated the role of H.E. mycelium in
modulating and reducing the loss of dopaminergic neurons. In the PC12
cell viability experiment, H.E. mycelium exhibited positive protective
eect to dopamine neuron in mice which represents the similar
neuroprotective properties in dopamine neurons to human PD [26,27].
Numerous studies have shown that MPP+-induces neurotoxicity in
cell and zebrash [28], and induces loss of dopaminergic neurons [29-
31]. Administration of MPP+ (1 mM) induced cell death in PC12 cells
is in consistent with several previous studies. MPP+-induced loss of
dopaminergic neurons indicates the selective damage to dopamine
neurons in cells. H. erinaceus extract promoted NGF synthesis by
hericenones from fruit bodies (Hericenone C-H) and by erinacines
from mycelium (erinacines A-I) [15]. H.E. mycelium reduced MPP+-
induced PC12 cell toxicity in a concentration dependent manner,
indicating that H.E. mycelium could contribute to the protective eect
Figure 4: The biosynthec pathway of dopamine neurotransmiers.
Tyrosine hydroxylase is the rate liming enzyme of the pathway.
Phenylalanine hydroxylase converts phenylalanine to tyrosine, tyrosine
hydroxylase hydroxylates tyrosine to L-DOPA. DOPA is converted to
dopamine by decarboxylase. Dopamine converts to Norepinephrine
by dopamine-β-hydroxylase and norepinephrine to epinephrine by
phenylethanolamine N- methyl transferase.
Figure 5: H.E. mycelium prevented MPTP-induced death of TH (Tyrosine
Hydroxylase)-posive neuron in striatum in mice.
(A). TH expression in striatum was assessed by immunohistochemistry.
(B). Quantave analysis of TH-posive cells in the striatum of
mouse brain. Dopamine neurons from mice are resistant to MPTP
neurotoxicity. There is signicant dierence in the average number
of TH posive neurons between the striatum of 0.3, 1 g/kg and that
of MPTP group. Data are presented as means ± SEM (n=10-15). One
way ANOVA and Newman-Keuls tests were used to analyze the data.
**P<0.01; ***p<0.001 when compared to control; ###P<0.001 when
compared to MPTP.
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Citaon: Yang PP, Lin CY, Lin TY, Chiang WC (2020) Hericium erinaceus Mycelium Exerts Neuroprotecve Eect in Parkinson’s Disease-in vitro
and in vivo Models. J Drug Res Dev 6(1): dx.doi.org/10.16966/2470-1009.150 5
Journal of Drug Research and Development
Open Access Journal
of dopaminergic neuron in PC12 cells. In 2002, Park YS, et al. [32]
reported that H. E. enhanced the synthesis of NGF (Nerve Growth
Factor) and BDNF (Brain-Derived Nerve Factor) in PC12 cells. BDNF
and NGF can be strongly expressed by dopaminergic neurons in
SNpc. But, in human and animal models of Parkinson’s disease, the
expression of NGF and BDNF levels are decreased [33,34]. us, NGF
level corresponds very well to the severity of PD. By summing up all
the evidence, a theory can be formed, in which erinacines of H.E.
mycelium upon ingested are transported to the brain where signicant
amount of NGF molecules are formed. e NGF molecules then
promote brain nerves in substantia nigra to grow where dopaminergic
neuron produces dopamine to a level that alleviates the Parkinson’s
disease.
In terms of eective dosage of H.E. mycelium, it was shown in
this study that it performed much better than Ganoderma lucidum.
Although it has been reported that Ganoderma lucidum has abilities
to induce neuronal dierentiation and prevent NGF-dependent PC12
neuronal cells from apoptosis [35], H.E. mycelium was more eective
in neuroprotection than Ganoderma lucidum in this PC12 cells study.
It may be due to that fact that a new cell-wall breaking technology
was employed and also ner particles were produced. It improves
the release of active ingredients from smaller H.E. mycelium particle
powders.
By using DPPH radical scavenging assay, widely used method to
evaluate the ability of mushroom to scavenge free radicals generated
from DPPH reagent [36,37], the present data also showed that H.E.
mycelium had more eective in scavenging DPPH radicals when
compared to Ganoderma lucidum. e radical inhibition eect of
Ganoderma lucidum was 59.3% at concentration of 1000 µg/ml, when
compared with H. E. mycelium’s 96.1% and with ascorbic acid’s 100%
at the same concentration.
Administration of MPTP was known to decrease the activity of
neuron and the density of TH-positive neurons, indicating that the
degeneration of the dopaminergic neurons in SNpc [38,39]. In our
present study, injection of MPTP into C57BL/6 Narl mice induced
signicant reduction of dopamine level and TH-positive area in the
striatum, suggesting that MPTP initially aects the dopaminergic
cell body in the substantia nigra pars compacta (SNpc) and then the
striatum where dopaminergic cells exert it’s function of release of
dopamine. Tyrosine hydroxylase (TH) is the initial enzyme in the
catecholamine synthesis pathway [40] and dopamine biosynthesis in
the central nervous system [41]. TH is activated to form more DOPA
and then dopamine by decarboxylation which is transferred into the
synaptic vesicle by the vesicular monoamine transporter (VMAT). In
addition, the loss of TH activity or expression is thought to contribute
to dopamine deciency, which is the most prominent at media level
of SNpc (substantia nigra pars compacta) [39]. In our current study,
the level of dopamine concentration and immunohistochemistry for
TH positive neurons revealed that the loss of dopamine neuron in PD
mice was dramatically prevented aer treatment of H.E. mycelium.
In consistent with other research, the results of our present study
showed that the immuno reactivity of TH was signicantly decreased
in MPTP-treated mice, suggesting that the majority of dopaminergic
neurons were lost in the Parkinson’s disease mouse model [42]. e
death of dopaminergic neurons led to decrease dopamine level in
the substantia nigra. e expression of TH positive neuron and
concentration of dopamine level were increased with the concentration
of H.E. mycelium from 0.3 g/kg to 1 g/kg in MPTP-induced mice. is
evidence suggested that the neuroprotection and anti-oxidation is involved
in the protective eect of H.E. mycelium on dopaminergic neurons.
Conclusion
In conclusion, we have used cell culture and animal models to
demonstrate the neuroprotective eect of H.E. myceliumin the PD
model. Our results indicate that H.E. mycelium increased the cell
viability of the MPP+ treated cells and performed in a very eective
antioxidative eect in vitro cell culture model using PC 12 cells. In vivo ,
H.E. mycelium exerts signicant neuroprotective eect by increasing
the dopamine level and the activity of tyrosine hydroxylase (TH) in
mice striatum. To sum up, our results suggest that H.E. mycelium
performs signicant protection of dopaminergic neuron under severe
conditions and is very eective in the treatment of damaged neuron in
the brain to recover in the case of Parkinson’s disease.
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