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Lion's Mane, Hericium erinaceus and Tiger Milk, Lignosus rhinocerotis (Higher Basidiomycetes) Medicinal Mushrooms Stimulate Neurite Outgrowth in Dissociated Cells of Brain, Spinal Cord, and Retina: An In Vitro Study

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Neurodegenerative disease is defined as a deterioration of the nervous system in the intellectual and cognitive capabilities. Statistics show that more than 80−90 million individuals age 65 and above in 2050 may be affected by neurodegenerative conditions like Alzheimer's and Parkinson's disease. Studies have shown that out of 2000 different types of edible and/or medicinal mushrooms, only a few countable mushrooms have been selected until now for neurohealth activity. Hericium erinaceus is one of the well-established medicinal mushrooms for neuronal health. It has been documented for its regenerative capability in peripheral nerve. Another mushroom used as traditional medicine is Lignosus rhinocerotis, which has been used for various illnesses. It has been documented for its neurite outgrowth potential in PC12 cells. Based on the regenerative capabilities of both the mushrooms, priority was given to select them for our study. The aim of this study was to investigate the potential of H. erinaceus and L. rhinocerotis to stimulate neurite outgrowth in dissociated cells of brain, spinal cord, and retina from chick embryo when compared to brain derived neurotrophic factor (BDNF). Neurite outgrowth activity was confirmed by the immu-nofluorescence method in all tissue samples. Treatment with different concentrations of extracts resulted in neuronal differentiation and neuronal elongation. H. erinaceus extract at 50 µg/mL triggered neurite outgrowth at 20.47%, 22.47%, and 21.70% in brain, spinal cord, and retinal cells. L. rhinocerotis sclerotium extract at 50 µg/mL induced maximum neurite outgrowth of 20.77% and 24.73% in brain and spinal cord, whereas 20.77% of neurite outgrowth was observed in retinal cells at 25 µg/mL, respectively.
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International Journal of Medicinal Mushrooms, 17(X): XXXXXX (2015)
Lion’s
Mane,
Hericium erinaceus
and
Tiger Milk,
Lignosus
rhinocerotis
(Higher
Basidiomycetes)
Medicinal Mushrooms Stimulate
Neurite
Outgrowth
in
Dissociated
Cells of Brain, Spinal Cord,
and Retina:
An In Vitro
Study
Snehlata Samberkar,1,2,* Sivasangkary Gandhi,1,3 Murali Naidu,1,2 Kah-Hui Wong,1,2
Jegadeesh Raman,1,3 & Vikineswary Sabaratnam1,3
¹Mushroom Research Centre, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur,
Malaysia; 2Department of Anatomy, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia; 3Institute of
Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
* Address all correspondence to: Snehlata P. Samberkar, Department of Anatomy, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur,
Malaysia; Tel: +60379673792; Fax: +603 79674724; E-mail: snehlata@um.edu.my
ABSTRACT: Neurodegenerative disease is defined as a deterioration of the nervous system in the intellectual and
cognitive capabilities. Statistics show that more than 8090 million individuals age 65 and above in 2050 may be
affected by neurodegenerative conditions like Alzheimers and Parkinsons disease. Studies have shown that out of
2000 different types of edible and/or medicinal mushrooms, only a few countable mushrooms have been selected
until now for neurohealth activity. Hericium erinaceus is one of the well-established medicinal mushrooms for neu-
ronal health. It has been documented for its regenerative capability in peripheral nerve. Another mushroom used as
traditional medicine is Lignosus rhinocerotis, which has been used for various illnesses. It has been documented for
its neurite outgrowth potential in PC12 cells. Based on the regenerative capabilities of both the mushrooms, prior-
ity was given to select them for our study. The aim of this study was to investigate the potential of H. erinaceus and
L. rhinocerotis to stimulate neurite outgrowth in dissociated cells of brain, spinal cord, and retina from chick embryo
when compared to brain derived neurotrophic factor (BDNF). Neurite outgrowth activity was
confirmed
by the immu-
nofluorescence method in all tissue samples. Treatment with different concentrations of extracts resulted in neuronal
differentiation and neuronal elongation. H. erinaceus extract at 50 µg/mL triggered neurite outgrowth at 20.47%,
22.47%, and 21.70% in brain, spinal cord, and retinal cells. L. rhinocerotis sclerotium extract at 50 µg/mL induced
maximum neurite outgrowth of 20.77% and 24.73% in brain and spinal cord, whereas 20.77% of neurite outgrowth
was observed in retinal cells at 25µg/mL, respectively.
KEY WORDS: medicinal mushroom, Lignosus rhinocerotis, Hericium erinaceus, neurite outgrowth activity, neuro-
degenerative disease
ABBREVIATIONS: BDNF, brain=derived neurotrophic factor; DAPI, 4′,6-diamidino-2-phenylindole; DMEM,
Dulbeccos modified Eagles medium; FITC, fluorescein isothiocyanate; PBS, phosphate-buffered saline.
I.
INTRODUCTION
Aging is an inevitable process and the fraction of
worlds population over 65 years of age is expected
to increase to 8090 million by the end of 2050.1
Out of many diseases that threaten aging humans,
neurodegenerative diseases such as Alzheimer s
disease, dementia, and Parkinsons disease2 and
retinal diseases like diabetic retinopathy, glaucoma,
and age-related macular degeneration can be very
traumatic.3 Neurohealth is a major concern as one
ages. Retrospective studies have proved that natural
products like mushrooms are suitable candidates for
neurohealth.4 Out of 14,000 species of familiar mush-
rooms,4 roughly 20004 are acknowledged to be
fit
for
human consumption. Researchers have reported that
1045-4403/15/$35.00 © 2015 Begell House, Inc. www.begellhouse.com 1
2
Samberkar et al.
many medicinal mushrooms promote neurotrophic
properties such as neurite outgrowth stimulation,5,6
nerve regeneration,7 neuroprotection,8 and anti-
oxidation.9 Mushrooms may have potential in the
prevention or treatment of age-related neurodegen-
erative complaints.10
The mushrooms selected for this study were
Hericium erinaceus (Bull.:Fr.)Pers. (Hericiaceae,
higher Basidiomycetes) and Lignosus rhinoc-
erotis (Cooke) Rivarden (Polyporaceae, higher
Basidiomycetes). H. erinaceus, a rare mushroom
also known as lions mane, monkeys head, and
Yamabu shitake, is scattered throughout North
America, Europe, and Asia.11,12 It has been used
in Chinese and Japanese cuisine and as an herbal
remedy to treat various human diseases including
gastric ulcers for hundreds of years.12 A health syrup
called “Houtou” is prepared from dried fruit bod-
ies. H. erinaceus tablets are used to treat ulcers,
inflammation,
and tumors of the alimentary canal.13
It is also evident that H. erinaceus has facilitated
functional recovery subsequent to peripheral nerve
injury.7 Based on this study, our focus is on regen-
eration of the central nervous system (CNS) using
brain, spinal cord, and retinal explants and cells after
dissociation, using chick embryo as a model.
L. rhinocerotis is also known as tigers milk
mushroom or “cendawan susu rimau” in the local
language. This species is distributed only in the tropi-
cal rainforest in regions of South China, Thailand,
Indonesia, Malaysia, Philippines, and Papua New
Guinea. L. rhinocerotis has been used as a general
tonic, antipyretic, and antipruritic; it has also been
used to treat fever, cancer, food poisoning, swollen
breasts, cough, and asthma and to assist in wound
healing, among other uses.14 Its usage is limited in
spite of its medicinal properties due to unavailability.
Sclerotium of L. rhinocerotis documented enhance-
ment of neurite outgrowth activity in PC12 cells.15
Based on their potential, in this study, both mush-
rooms with different concentrations (25100 µg/mL)
were treated with tissue samples and compared with
negative and positive controls. Neurite extensions
were confirmed by immunofluorescence staining.
II. MATERIALS
AND
METHODS
A.
Preparation
of
Mushroom
Aqueous Extracts
Mushrooms require the correct combination of
humidity, temperature, substrate (growth medium),
and inoculum (spawn) to grow. In Malaysia,
H. erinaceus is cultivated on a substrate containing
rubberwood sawdust, rice bran, and calcium carbon-
ate at a ratio of 100:5:1. After 2 months of spawn run
at 27±2 to 32±2°C at a mushroom farm (Ganofarm
Ltd., Tanjung Sepat, Selangor, Malaysia), approxi-
mately 300 g of fresh fruit body per 800 g of substrate
was harvested.16 Fresh fruiting bodies of H. erinaceus
were purchased from the mushroom farm. Fresh
fruit bodies were sliced, frozen, and freeze-dried.
The freeze-dried fruit bodies were then blended in
a Waring commercial blender and stored in airtight
containers at C prior to assay.16 L. rhinocerotis,
a rare species, is found in the forests of Malaysia.
In this study, the freeze-dried powder of sclerotia
of cultivated L. rhinocerotis10,17 was purchased
from Ligno Biotek Sdn Bhd (batch no. TM02). The
freeze-dried powders of both mushrooms were then
soaked separately in distilled water (1:20, w/v) and
were agitated at 150 rpm for 24 h.15 The mixture
was then double boiled in a water bath at 100°C
for 30 min, cooled, and filtered by Whatman filter
paper No. 4. The aqueous extract was freeze-dried
and kept at 20°C prior to use.6,18
B.
Preparation
of Explants
Culture
and Trypsinization
Fertilized chicken eggs were collected from Charoen
Pokphand Jaya Farm (M) Sdn Bhd (Negeri Sembilan,
Malaysia) and then were incubated at 39°C in a
humidified
incubator. The brain (day 4), spinal cord
(day 6), and retina (day 9) were dissected on their
respective days.19 Based on a modified method of
Gibco Life Technologies (Selangor, Malaysia), the
tissue samples (brain, spinal cord, and retina) were
finely chopped, washed with phosphate-buffered
saline (PBS) twice, and centrifuged at 3000 rpm
for 3 min after adding trypsin. The supernatant
International Journal of Medicinal Mushrooms
Medicinal Mushrooms Induce Neurite Outgrowth in Brain, Spinal Cord, and Retinal Cells
3
2
2
was discarded and the cell pellet was resuspended
twice with 25 mL of prewarmed (37°C) complete
media. The samples were centrifuged at 3000 rpm
for 35 min. The supernatant was discarded and
the cell pellet was added to fresh complete media
dilution in blocking buffer) for 1 h. The cells were
washed and then incubated with secondary antibody,
anti-rabbit IgG-fluorescein isothiocyanate (FITC)
antibody produced in sheep (Sigma, St. Louis, MO,
USA) (1:80 dilution in blocking buffer) for 1 h in
and incubated at 37±2°C in a 5% CO
incubator for 24 h.
C. Neurite
Outgrowth
Assay
humidified the dark at room temperature. The cells were then
washed thrice. The coverslips were then mounted
with 4′,6-diamidino-2-phenylindole (DAPI), which
stained the nucleus. Images were observed and cap-
tured with a
fluorescent
microscope (Nikon Eclipse
Two-day-old cultured cells were seeded into 12-well
plates at a cell density of 5×104 cells per well. The
mushroom aqueous extracts in Dulbeccos modified
Eagles medium (DMEM) at concentrations of 25,
50, 75, and 100 µg/mL (w/v), and brain-derived
neurotrophic factor (BDNF) at 10 ng/mL (w/v) were
tested for neurite outgrowth stimulation activity.
Cells in complete DMEM without treatment served
as the negative control. Plates were incubated at
80i microscope using FITC and DAPI filters).
F. Statistical
Analysis
All experiments were carried out in three repli-
cates. Results were expressed as the means ± SD.
All data were subjected to analysis of variance
using GraphPad Prism Statistical Software version
7 (GraphPad Software Inc., La Jolla, CA, USA). The
37±2°C in a 5% CO incubator for 2 days.15 differences among samples were evaluated using
Duncans multiple range test, where p<0.05 was
D.
Scoring
of
Neurites
Neurite extensions were scored under an inverted
microscope (Nikon Eclipse TS100) with the aid of
a handheld counter. A cell was scored positive for
bearing neurites if it had at least one thin extension
longer than the diameter of its cell body.20 In a well,
10 fields with an average of randomly chosen 250
300 cells per well were examined and photographed
using a Nikon DS-Fi1 camera and were processed
with Nikons NIS-Elements D imaging software.6
E.
Neurofilament Staining
A
neurofilament
was used as an indicator for neurite
outgrowth and immunofluorescence staining was
used for confirming neuronal extension, which is
an increase in axonal length. Based on the standard
method,21 primary neuronal cells were seeded in
12-well plates and exposed to treatment for 2 days.
The cells were fixed with 4% paraformaldehyde
for 20 min at room temperature. After two washes
with PBS, the cells were incubated with primary
antibody,
antineurofilament
200 antibody produced
in rabbits (Sigma, St. Louis, MO, USA) (1:80
considered significant.
III. RESULTS
AND
DISCUSSION
Neurons in the mature CNS are unable to regenerate
injured axons and the neurons that remain unin-
jured are unable to form novel connections that
might compensate for ones that have been lost.22
Subsequently, due to a break in the communication
between healthy neurons, a cascade of events takes
place that leads to neuronal degeneration and cell
death. The factors responsible for failure of regen-
eration are several and include poor regenerative
ability of CNS neurons, inhibitory properties of
astrocytes,23 and inhibitory molecules produced
by oligodendrocytes and myelin.24 Overpowering
these issues will facilitate the nerve regeneration for
restoration of function following damage, through
accident, injury, or neurodegenerative disease.
Natural products have been traditionally
accepted as remedies due to the popular belief that
they present minor side effects.25 In traditional
Chinese medicine, mushrooms have always been
prepared for medicinal use by hot water extraction.
The number of mushrooms, however, studied for
Volume 17, Number X, 2015
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Samberkar et al.
FIG. 1: Effect of varying concentrations of H. erinaceus and Lignosus rhinocerotis aqueous extracts on neurite out-
growth stimulation on chick embryos brain cells.
neurohealth activity are few and H. erinaceus10 is
one of them. Its chemical composition was docu-
mented by Kawagishi et
al.
5
Further study on aqueous
extracts proved that polysaccharides could induce
neuronal differentiation and promote neuronal sur-
vival.26 Ongoing research in our laboratory shows
that H. erinaceus,6 L. rhinocerotis,15,27 and Pleurotus
giganteus (Berk.) Karunarathna & K.D. Hyde
(morning glory mushroom, cows stomach mush-
room)28 exhibit neurite outgrowth stimulatory effects
in NG108-15 and PC12 cell lines. Inspired by this,
researchers focused their goal on peripheral nerve
regeneration following crush injury to the rat pero-
neal nerve by using aqueous extract of H. erinaceus.
This study proved that daily administration of aque-
ous extract of H. erinaceus has
beneficial
effects on
recovery of the injured rat peroneal nerve in early
stages of regeneration.7
Another medicinal mushroom is L. rhinocerotis.
Retrospective studies have shown that an aqueous
extract of L. rhinocerotis sclerotium induced neu-
rite outgrowths of 24.4% and 42.1% at 20 µg/mL
(w/v) of aqueous extract alone and a combination of
20 µg/mL (w/v) aqueous extract and 30 ng/mL (w/v)
of NGF, respectively, in rat pheochromocytoma cells
(PC12 cells).15 Similarly, the present study showed
neurite outgrowth of 20.77% and 24.73% at 50 µg/mL
in brain and spinal cord cells and 20.77% at 25 µg/mL
in retinal cells. Consistent with our previous study,
the sclerotia of L. rhinocerotis reported neurite
outgrowth in N2a.29 It was recently shown that scle-
rotial extract performs better than mycelial extract.29
Advanced study shows maximum neurite extension
for L. rhinocerotis and curcumin at 21.1% at 20
µg/mL and 29.47% at 10 µg/mL on PC-12 cells.30
Combining 20 µg/mL of L. rhinocerotis with 1 µg/
mL curcumin gave 27.2% neurite extension in PC12
cells.30 Taken as a whole, these medicinal mushrooms
have shown neurological properties such as neuronal
survival and neurite outgrowth activities including
improvement in recovery and function in both in vivo
and in vitro mammalian nervous systems.31
The aqueous extracts of H. erinaceus and L. rhi-
nocerotis showed a gradual dose-dependent twofold
increase in neurite outgrowth stimulation at 25 and
50 µg/mL concentration compared to the negative
control. Figure 1 shows the neurite outgrowth stimu-
lation on brain cells after 48 h incubation decreased
the extension in a dose-dependent manner observed
at 75 and 100 µg/mL concentrations. The maximal
stimulated outgrowth on brain cells treated with
aqueous extracts of H. erinaceus and L. rhinocerotis
was 20.47% and 20.77%, respectively, at 50 µg/mL,
comparable to that of the BDNF-treated cells
(positive control), whereas H. erinaceus showed
significant (p<0.05) neurite outgrowth of 18.73%
International Journal of Medicinal Mushrooms
Medicinal Mushrooms Induce Neurite Outgrowth in Brain, Spinal Cord, and Retinal Cells
5
FIG. 2: Effects of varying concentrations of H. erinaceus and Lingnosus rhinocerotis aqueous extract on in vitro neu-
rite outgrowth stimulation on chick embryos spinal cells.
at a concentration of 25 µg/mL. H. erinaceus and
L. rhinocerotis aqueous extracts showed signifi-
cant (p<0.05) neurite outgrowth of 22.47% and
24.73%, respectively, at 50 µg/mL on the spinal
cord (Fig. 2). The aqueous extract of L. rhinocero-
tis showed significant (p<0.05) neurite outgrowth
(20.77%) at 25 µg/mL on retinal cells; in contrast,
the aqueous extract of H. erinaceus (21.70%)
exerted its maximum neurite growth at 50 µg/mL
(Fig. 3). Comparing both of the aqueous extracts,
H. erinaceus exerted a significantly potent neurite
outgrowth on retinal cells at a lower concentra-
tion compared to L. rhinocerotis aqueous extract.
Retinal cell stimulation was significant (p<0.05)
at 50 µg/mL by H. erinaceus. Neurite outgrowth
is confirmed by neurofilament staining as shown
in Fig. 4. Figures 4A, D, and G show BDNF at
the concentration of 10 ng/mL used as a positive
control in the brain, spinal cord, and retina. Figures
4B, E, and H show the negative control in all three
FIG. 3: Effects of varying concentrations of H. erinaceus and Lignosus rhinocerotis aqueous extract on in vitro neu-
rite outgrowth stimulation on chick embryos retina cells.
Volume 17, Number X, 2015
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Samberkar et al.
FIG. 4: Fluorescent microscopy image of in vitro neurite outgrowth in chicken embryos brain, spinal cord. and retinal
cells by Lignosus rhinocerotis aqueous extracts. Brain and spinal cord cells were treated with hot aqueous extract
L. rhinocerotis at a concentration of 50 and 25 µg/mL for retinal cells after 48 h of incubation at 37±2°C in a 5% CO
2
humidified incubator. (A) Brain positive control: BDNF(10 ng/mL). (B) Negative control of brain cells without extract.
(C) Brain cells treated with hot aqueous extract of L. rhinocerotis at 50 µg/mL. (D) Spinal cord positive control: BDNF
(10 ng/mL). (E) Negative control of spinal cord cells without extract. (F) Spinal cord cells treated with hot extract of L.
rhinocerotis at 50 µg/mL. (G) Retina positive control: BDNF (10 ng/mL). (H) Negative control of retinal cells without
extract. (I) Retinal cells treated with hot aqueous extract of L. rhinocerotis at 25 µg/mL. (C), (F), and (I) Cells show
an exuberant long neurite outgrowth (arrow) as compared to (A) (D), and (G).
samples, which have either no neurite, or neurite
with insufficient length to be scored as positive.
Figures 4C, F, and I show neurite outgrowth in the
brain, spinal cord, and retinal cells confirmed by
neurofilament
staining. Neurite extension is marked
by an arrow. Recent research suggested that these
neurofilaments are closely related to many neuro-
degenerative diseases, such as amyotrophic lateral
sclerosis, Parkinsons diseae, and Alzheimers dis-
ease. Using in vitro assays, cultures, and transgenic
mice, these studies provided new insights into neuro-
filament function. The function of each subunit,
the relationship of neurofilaments with other cyto-
skeletal elements and their clinical significance are
topics of increasing attention.
IV.
CONCLUSIONS
H. erinaceus and L. rhinocerotis aqueous extracts
were examined for neurite outgrowth activity in
International Journal of Medicinal Mushrooms
Medicinal Mushrooms Induce Neurite Outgrowth in Brain, Spinal Cord, and Retinal Cells
7
the brain, spinal cord, and retinal cells of chicken
embryo. Four different concentrations (25, 50, 75,
and 100 µg/mL) were tested. Among the concen-
trations, H. erinaceus aqueous extract, 50 µg/mL,
stimulated neurite outgrowth in brain and spi-
nal cord cells. L. rhinocerotis sclerotium extract
showed similar neurite outgrowth activity in reti-
nal cells at 25 µg/mL after 48 h of incubation.
Immunofluorescence staining confirmed neurite
outgrowth.
ACKNOWLEDGMENTS
The authors thank the University of Malaya for
Bantuan Kecil Penyelidikan (BKP) grant BK003-
013, RG 193/11AFR, and UMRG RP 005B-13
AFR, and Mrs. Cheng Poh Guat of Vita Agrotech,
a mushroom farm in Tanjung Sepat, Selangor,
Malaysia for the continuous supply of fresh
H. erinaceus mushroom. The technical assistance
by the staff of the Department of Anatomy is greatly
appreciated.
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International Journal of Medicinal Mushrooms
... Lion's many also appear to help with memory. One of the systemic reviews found that mushroom extracts like that of Lion's Mane have a positive impact on cognition [12][13][14][15]. ...
... In experimental studies, Lion's mane extract appears to promote nerve recovery and growth. Thus, it may have a role as a health supplement in preventing neuropathies [13,16,17]. ...
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Lion's Mane, a distinctive mushroom found across the Northern Hemisphere, has captivated the world with its unique appearance and potential health benefits. This article explores the diverse facets of Lion's Mane, from its taxonomy and habitat to its rich nutritional composition, including proteins, carbohydrates, fats, vitamins, minerals, and an array of biologically active compounds, notably polysaccharides and phenolic compounds. Lion's Mane is emerging as a potent dietary supplement, with research highlighting its positive impact on brain health, nerve recovery, mood stabilization, gastric health, heart health, blood sugar regulation, cancer prevention, and immunity enhancement. While Lion's Mane offers promising health benefits, it is essential to recognize its role as a complementary dietary addition rather than a substitute for medical treatments. Fortunately, Lion's Mane is generally considered safe for prolonged consumption, with minimal reported side effects. As research continues to unveil the multifaceted potential of the Lion's Mane, it stands as a fascinating and versatile natural resource with the promise of contributing to improved well-being and longevity.
... TMP thus promoted the regeneration of neuronal cells in vivo and in vitro, and significantly inhibited neuroinflammation allowing improved clinical function in mouse models. Samberkar and colleagues [71] found the mushrooms Hericium erinaceus (Lion's Mane) and Lignosus rhinocerotis (Tiger milk) stimulated neurite outgrowth in dissociated cells of brain and spinal cord from chick embryos. These effects were dose dependent. ...
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Spinal cord injury (SCI) affects approximately 40 million people each year worldwide and no effective medicine has been found to ameliorate the disabling effects of SCI. Psychoplastogens are a heterogenous group of medicines defined by their function rather than their chemical structure. These medicines catalyze the growth of new neurons (viz. neurogenesis) and stimulate the formation of new connections between neurons (viz. synaptogenesis). Examples of psychoplastogens include ketamine, psilocybin, and lysergic acid diethylamide (LSD). Several psychoplastogens exhibit promise as potential therapeutic agents in the treatment of SCI. In pre-clinical studies, the psychoplastogen LSD aided in recovery from SCI. An anecdotal report suggests the psychoplastogen psilocybin assisted a paralyzed patient recover from SCI. Finally, other mycological psychoplastogens have also shown promise as treatments for SCI including Tricholoma matsutake (Pine mushroom), Hericium erinaceus (Lion’s Mane), and Lignosus rhinocerotis (Tiger milk). Racemic ketamine is an inexpensive general anesthetic, a rapid-acting antidepressant medication, and a psychoplastogen. This medication, which is safe when used in low doses, stimulates the growth of nervous system structures, thus making it a potential option for individuals who suffer from SCI. Based upon a review of the existing literature regarding the aforementioned psychoplastogens, we hypothesize that daily low dose racemic ketamine may assist in recovery from SCI. We recommend clinical trials to determine the efficacy, dosage range, and optimal dosing frequency for the treatment of SCI.
... It has been used in TCM practices for thousands of years to alleviate gastrointestinal discomfort. Therapeutic potentials of H. erinaceus have been demonstrated in preclinical studies of neuroregeneration [120][121][122], depressive-like disorder [78,123,124], cerebellar ataxia [125][126][127], PD [128], AD [129,130], and traumatic brain injury [131]. A total of 80 secondary metabolites, namely hericenones, erinacines [132,133], erinapyrones [134,135], sterols, fatty acids, and fumitremorgin C [136] have been isolated from the fruiting bodies and mycelium of mushrooms. ...
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Spinocerebellar ataxia type 3 (SCA3) is an autosomal dominant neurodegenerative disorder that gives rise to motor incoordination and progressive functional disabilities. Although pharmacological interventions have revealed promising prospects in the management of SCA3, adverse effects may become unbearable. The use of herbal remedies in traditional Chinese medicine (TCM) may serve as potential alternative medicines to delay the progression of the disease. This systematic review is intended to identify, appraise, and summarize the findings of studies pertaining to the therapeutic roles of herbal remedies in TCM targeting oxidative stress in the management of SCA3. A literature search for relevant articles published from 1 January 2013 to 30 June 2023 in three databases, namely PubMed, Web of Science, and Scopus, was carried out according to the procedures of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). A total of ten preclinical studies met the inclusion criteria of the systematic review. We recognized the therapeutic potential of Brassica napus, Codonopsis pilosula, Curcuma sp., Gardenia jasminoides, Gastrodia elata, Ginkgo biloba, Glycyrrhiza inflata, Hericium erinaceus, Hyptis sp., Paeonia lactiflora, Panax ginseng, Poria cocos, Pueraria lobata, Rehmannia glutinosa, and Scrophularia ningpoensis. We identified the types of preclinical models expressing polyglutamine (polyQ) expanded mutant protein (mATXN3), inducers of oxidative stress that mimic the SCA3 pathogenesis, and effective doses of the herbal remedies. The modes of action contributing to the attenuation of oxidative stress are activation of antioxidant pathways, ubiquitin–proteasome system and autophagy, regulation of apoptosis, proinflammatory signaling pathway and chaperones, regulation of mitochondrial function and biogenesis, and restoration of neurotransmission and synaptic plasticity. In conclusion, herbal remedies in TCM may possibly delay the progression of SCA3, therefore providing justification for clinical trials.
... Lion's Mane extract can induce nerve growth factor (NGF) synthesis in human astrocytoma cells; the mice fed with 5% Lion's Mane powder for 7 days had increased NGF mRNA expression in their hippocampus (15). The enhanced neurotrophin expression by Lion's Mane was also confirmed in studies using different glioma cell lines (16)(17)(18). Another study showed that when administered to mice that recently had a crush-injury to their hind limb, Lion's Mane accelerated the functional recovery of the limb. This most likely occurred by promoting peripheral nerve axon regeneration and reinnervation and activation of neurotrophic signaling at the ipsilateral dorsal root ganglia (19). ...
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Lion's Mane (Hericium erinaceus) has historically been used as traditional medicine in Asia and Europe for its potential benefits in fighting infection and cancer. It has gained interest in the neurodegenerative disease field because of its mechanisms of action; these include anti-inflammation, neuroprotection, and promoting neurite growth demonstrated in various cell and animal models. A very small, double-blind, placebo-controlled trial in patients with mild cognitive impairment showed a temporary improvement in cognitive function; this finding has yet to be replicated. However, there have been no studies in ALS cell or animal models or in humans with ALS. Lion's Mane appears safe and inexpensive when consumed in powder or capsule, but one anaphylactic case was reported after a patient consumed fresh Lion's Mane mushroom. Currently, we do not have enough information to support the use of Lion's Mane for treating ALS. We support further research in ALS disease models and clinical trials to study its efficacy.
... Experimental evidences and mechanism of action H. erinaceus significantly influences the Norepinephrine system's initiation and regulation of neurotrophins. H. erinaceus active ingredients, particularly the erinacines and hericenones, have potentive nerve growth factor stimulating properties and exhibit impressive nerve outgrowth functions in a variety of cell lines as well as dissociated neurons within the brain, vertebral column, and retina [51][52][53]. A daily, eight-week oral supplement with H. erinaceus (80% mycelium extract and 20% fruiting body extract), together with a low-calorie diet plan, has been shown to reduce sadness, stress, insomnia, and food addiction, according to a recent study on 77 subjects who were overweight or obese, eating, in contrast to participants on a reduced diet alone. ...
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Background One of the most important healthcare challenges in the world today is neurological disorders. Pose lifestyle changes are linked to a significantly higher risk of chronic illnesses and diseases, placing a significant financial and healthcare burden on society at large. In this review article, we focused on the various neuro-nutraceutical herbs and their beneficial roles in neurodegenerative disorders. Main body of the abstract An extensive literature review was done from the standard databases such as Scopus, Elsevier, and PubMed using standard keywords “Nutraceuticals”, “Neuro-nutraceuticals”, “Neurodegenerative disorders”. Numerous "neuro-nutraceuticals" are natural plant compounds with dietary and pharmaceutical components that are intended to improve cerebral blood flow along with illness prevention and control. These compounds are found in food, herbal medicines, and nutritional supplements such as Bacopa monnieri, Curcuma longa, Asparagus racemosus, Helicteres angustifolia, Hericium erinaceus, Crocus sativus, Uncaria tomentosa, Centella asiatica, Convolvulus pluricaulis, Moringa oleifera. Short conclusion While discussing the neuroprotective and the neuromodulatory properties of various neuro-nutraceuticals, we rationally postulate here their molecular mechanisms. Additionally, compared to single-target medicines, which may have unfavourable side effects, these herbs are believed to be safer and to provide a more holistic improvement in brain health. Graphical Abstract
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The burden of neurological disorders is growing substantially with limited therapeutic options, urging the consideration and assessment of alternative strategies. In this regard, we aimed to elucidate the phytochemical profile of the petroleum ether extract (PEE) of three palm tree species: Aiphanes eggersii Burret, Carpoxylon macrospermum H. Wendl. & Drude, and Jubaeopsis caffra Becc. (Family Arecaceae), and to evaluate their neuroprotective effect in monosodium glutamate (MSG)-induced excitotoxicity model for the first time. We identified a total of 48, 18, and 45 compounds in A. eggersii, C. macrospermum, and J. caffra, constituting 79.41 %, 60.45 %, and 76.35 % of the total detected compounds, respectively. A. eggersii extract was rich in the methyl esters of fatty acids (65.08 %) especially methyl dodecanoate (17.72 %). C. macrospermum was exclusively prolific by the triterpene 3β-methoxy-d:c-friedo-b’:a'-neogammacer-9(11)-ene (40.36 %), while J. caffra was noticeable by hydrocarbons (30.14 %) and lupeol derivatives (19.79 %). The biochemical and histopathological analysis showed that the tested extracts significantly reduced the oxidative stress, especially at the highest tested dose (1000 mg/kg). The extracts also reduced the activity of induced nitric oxide synthetase, Ca⁺² level, and NR2B subunit expression and attenuated apoptosis and DNA damage. The docking results show that most active natural compounds bind to SOD-1 and NR2B-NMDARs, verifying the credibility of the biological findings. To sum up, the PEE of the three investigated palm tree species possessed a unique blend of lipophilic bioactive constituents that exert promising neuroprotective potential against MSG-induced excitoneurotoxicity. However, further preclinical investigation and pharmaceutical formulation are needed.
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This article delves into the profound significance of the early diagnosis of Alzheimer’s disease (AD), the leading cause of dementia worldwide. With no current cure for AD, early detection stands as a cornerstone in managing the disease. Early diagnosis not only enables symptomatic treatment to enhance the quality of life but also facilitates proactive planning, addressing health care and living arrangements for the future. Additionally, early diagnosis can promote participation in clinical trials, granting patients access to emerging treatments. An essential component to this early detection is a robust understanding of the disease's causes. The paper examines the pathological indicators, such as beta-amyloid plaques and neurofibrillary tangles, while highlighting the multifaceted origins of AD encompassing genetics, environmental factors, inflammation, and potential links with other diseases. An in-depth discussion on the influence of the environment further illustrates the complex interplay between genetics and external factors. Toxic chemicals, lifestyle choices in western societies, and other environmental determinants are scrutinized for their potential role in AD onset. In summary, the piece underscores the importance of a holistic understanding of Alzheimer's etiology, emphasizing that only through comprehensive knowledge can we aspire to identify, manage, and ultimately find a cure for this debilitating condition.
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A novel diterpenoid, erinacine D, was isolated from the cultured mycelia of Hericium erinaceum. The structure of the compound was determined by interpretation of the spectral data and chemical reaction. This compound showed stimulating activity of nerve growth factor (NGF)-synthesis.
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Hericium erinaceus a culinary and medicinal mushroom is a well established candidate for brain and nerve health. Ganoderma lucidum, Grifola frondosa and Sarcodon scabrosus have been reported to have neurite outgrowth and neuronal health benefits. The number of mushrooms, however, studied for neurohealth activity are few compared to the more than 2 000 species of edible and / or medicinal mushrooms identified. In the on-going search for other potent culinary and / or medicinal mushrooms, indigenous mushrooms used in traditional medicines such as Lignosus rhinocerotis and Ganoderma neo-japonicum are also being investigated. Further, the edible mushroom, Pleurotus giganteus can be a potential candidate, too. Can these edible and medicinal mushrooms be tapped to tackle the health concerns of the aging population which is projected to be more than 80-90 million of people age 65 and above in 2050 who may be affected by age-related neurodegenerative disorders. Scientific validation is needed if these mushrooms are to be considered and this can be achieved by understanding the molecular and biochemical mechanisms involved in the stimulation of neurite outgrowth. Though it is difficult to extrapolate the in vitro studies to what may happen in the human brain, studies have shown that there can be improvement in cognitive abilities of the aged if the mushroom is incorporated in their daily diets.
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Abstract Mushrooms have long been used not only as food but also for the treatment of various ailments. Although at its infancy, accumulated evidence suggested that culinary-medicinal mushrooms may play an important role in the prevention of many age-associated neurological dysfunctions, including Alzheimer's and Parkinson's diseases. Therefore, efforts have been devoted to a search for more mushroom species that may improve memory and cognition functions. Such mushrooms include Hericium erinaceus, Ganoderma lucidum, Sarcodon spp., Antrodia camphorata, Pleurotus giganteus, Lignosus rhinocerotis, Grifola frondosa, and many more. Here, we review over 20 different brain-improving culinary-medicinal mushrooms and at least 80 different bioactive secondary metabolites isolated from them. The mushrooms (either extracts from basidiocarps/mycelia or isolated compounds) reduced beta amyloid-induced neurotoxicity and had anti-acetylcholinesterase, neurite outgrowth stimulation, nerve growth factor (NGF) synthesis, neuroprotective, antioxidant, and anti-(neuro)inflammatory effects. The in vitro and in vivo studies on the molecular mechanisms responsible for the bioactive effects of mushrooms are also discussed. Mushrooms can be considered as useful therapeutic agents in the management and/or treatment of neurodegeneration diseases. However, this review focuses on in vitro evidence and clinical trials with humans are needed.
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Mushrooms are not only regarded as gourmet cuisine but also as therapeutic agent to promote cognition health. However, little toxicological information is available regarding their safety. Therefore, the aim of this study was to screen selected ethno-pharmacologically important mushrooms for stimulatory effects on neurite outgrowth and to test for any cytotoxicity. The stimulatory effect of mushrooms on neurite outgrowth was assessed in differentiating mouse neuroblastoma (N2a) cells. Neurite length was measured using Image-Pro Insight processor system. Neuritogenesis activity was further validated by fluorescence immunocytochemical staining of neurofilaments. In vitro cytotoxicity was investigated by using mouse embryonic fibroblast (BALB/3T3) and N2a cells for any embryo- and neuro-toxic effects; respectively. Aqueous extracts of Ganoderma lucidum, Lignosus rhinocerotis, Pleurotus giganteus and Grifola frondosa; as well as an ethanol extract of Cordyceps militaris significantly (p < 0.05) promoted the neurite outgrowth in N2a cells by 38.4 +/- 4.2%, 38.1 +/- 2.6%, 33.4 +/- 4.6%, 33.7 +/- 1.5%, and 35.8 +/- 3.4%; respectively. The IC50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts. Our results indicate that G. lucidum, L. rhinocerotis, P. giganteus, G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health.
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Peripheral nerve injury represents a huge burden to society. Following peripheral nerve injury, improved behavioral outcome may be the most important evidence of functionality of axonal regeneration after any repair strategy. Nerve-crush injury is a well-established axonotmetic model in experimental regeneration studies to investigate the impact of various pharmacological treatments. Hericium erinaceus is a temperate mushroom but is now being cultivated in tropical Malaysia. In this study, we investigated the activity of aqueous extract of H. erinaceus fresh fruitbodies in promoting functional recovery following an axonotmetic peroneal nerve injury in adult female Sprague-Dawley rats with a long-term view toward the possible use of this mushroom in the treatment of nerve injury. Functional recovery was assessed in the behavioral experiment by walking-track analysis and toe-spreading refl ex. The peroneal functional index (PFI) was determined before surgery and after surgery , as the rats showed signs of recovery. Analysis of the PFI indicated that the return of hind-limb function occurred by day 10 to 14 and by day 14 to 17 in the treated and control (nontreated) groups, respectively. Normal toe-spreading in the crushed limb was achieved by day 7 to 10 and day 12 to 17 in the treated and control group, respectively. These results suggest that daily administration of aqueous extract of H. erinaceus fresh fruitbodies has a benefi cial effect on the recovery of injured rat peroneal nerve in the early stages of regeneration. The PFI and toe-spreading refl ex improved faster in the treated group than in the nontreated group.
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Lignosus rhinocerus ("Tiger's Milk" mushroom, or local Malay name "cendawan susu rimau (harimau)") is one of the most important traditional medicinal mushrooms found in Malaysia. The local communities have been using this mushroom to treat asthma, fever, cough, cancer, food poisoning, wound healing, as a tonic, as well as to improve immunity and maintain general health. In this study, five isolates of L. rhinocerus from three different geographical regions in Malaysia with distances of more than 100 km were collected, and their internal transcribed spacer (ITS) regions of ribosomal RNA were sequenced and compared. Although these isolates were sampled from different geographical locations, they shared high nucleotide identity in ITS regions. A specific primer was designed based on the DNA sequences within the ITS1 region and paired with ITS4 primer in order to use as specific PCR-based genetic marker for identifying L. rhinocerus.