Mushroom varieties – effects on health

Scientific Investigation from Mushrooms and Health

2008

Background
Mushrooms and Health 2008 covered Agaricus bisporus and 17 culinary
specialty and nutraceutical specialty mushrooms; however some
additional culinary and nutraceutical mushrooms have also been included
where significant information was available on consumption and health.
The scientific literature is categorised both by health condition and
subsequently, the information is also grouped by mushroom variety for
readers interested in specific mushroom varieties.

Agaricus bisporus (common white button,

brown/crimini, portabella)
It has been demonstrated that dietary supplementation with white button
mushrooms (Agaricus bisporus) enhances
natural killer (NK) cell activity in C57BL/6 mice,
suggesting that increased intake of white button
mushrooms may promote innate immunity
against tumours and viruses through the
enhancement of NK activity (Wu et al. 2007).

Extracts from Agaricus bisporus mushrooms

have been suggested as potential breast cancer
chemopreventive agents, as they suppress
aromatase activity and estrogen biosynthesis. A

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recent study has evaluated the activity of mushroom extracts in the
estrogen receptor-positive/aromatase-positive MCF-7aro cell line in vitro
and in vivo. Mushroom extract decreased testosterone-induced cell
proliferation in MCF-7aro cells but had no effect on MCF-10A, a non-
tumourigenic cell line. Most potent mushroom chemicals are soluble in
ethyl acetate. The major active compounds found in the ethyl acetate
fraction were unsaturated fatty acids such as linoleic acid, linolenic acid,
and conjugated linoleic acid. The interaction of linoleic acid and
conjugated linoleic acid with aromatase mutants expressed in Chinese
hamster ovary cells showed that these fatty acids inhibited aromatase
with similar potency and that mutations at the active site regions affect
its interaction with these two fatty acids. Whereas these results suggest
that these two compounds bind to the active site of aromatase, the
inhibition kinetic analysis indicated that they are non-competitive
inhibitors with respect to androstenedione. As only conjugated linoleic
acid was found to inhibit the testosterone-dependent proliferation of MCF-
7aro cells, the physiologically relevant aromatase inhibitors in
mushrooms are most likely conjugated linoleic acid and its derivatives.
The in vivo action of mushroom chemicals was shown using nude mice
injected with MCF-7aro cells. The studies showed that the mushroom
extract decreased both tumour cell proliferation and tumour weight with
no effect on the rate of apoptosis (Chen et al. 2006).

The edible mushroom lectin from Agaricus bisporus has been reported to
have anti-proliferative effects on a range of cell types. A study has been
undertaken to determine whether it might have inhibitory activity on
Tenon's capsule fibroblasts in in vitro models of wound healing and
therefore have a use in the modification of scar formation after glaucoma
surgery. Human ocular fibroblasts in monolayers and in three-
dimensional collagen lattices were exposed to Agaricus bisporus (0-100
mg/ml). Agaricus bisporus caused a dose-dependent inhibition of
proliferation and lattice contraction without significant toxicity. The data
showed that Agaricus bisporus possesses key features required of an
agent that might control scarring processes and suggest that Agaricus
bisporus may be especially useful where subtle modification of healing
may be needed, although further studies are required (Batterbury et al.
2002).

The effect on epithelial cells of a Gal beta-1,3-GalNAc-binding lectin, from

the edible mushroom Agaricus bisporus lectin (ABL) has been evaluated.
ABL (25mg/ml) inhibited incorporation of [3H]-thymidine into DNA of
HT29 colon cancer cells by 87%, Caco-2 colon cancer cells by 16%, MCF-
7 breast cancer cells by 50%, and Rama-27 rat mammary fibroblasts by
55% when these cells were grown for 24h in serum-free medium. When

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assessed by cell count, similar inhibition of proliferation of HT29 cells by
ABL was found. ABL (0.2mg/ml) caused no cytotoxicity to HT29, MCF-7,
and Rama-27 cells as measured by trypan blue exclusion, and inhibition
of proliferation in HT29 cells caused by 50mg/ml ABL was reversible after
removal of the lectin. A. bisporus lectin appears to be a reversible non-
cytotoxic inhibitor of epithelial cell proliferation which deserves further
study as a potential anti-cancer agent (Yu et al. 1993).

The reversibility of the anti-proliferative effect of Agaricus bisporus lectin

is associated with its release from cancer cells after internalization. The
internalization and subsequent slow release, with little degradation of the
lectin, reflects the tendency of lectins to resist biodegradation and implies
that other endogenous or exogenous lectins may be processed in this
way by intestinal epithelial cells (Yu et al. 2000).

The three-dimensional structure of the lectin from Agaricus bisporus has

been determined by x-ray diffraction. The protein is a tetramer with 222
symmetry, and each monomer presents a novel fold with two beta sheets
connected by a helix-loop-helix motif. The T-antigen disaccharide, Gal
beta 1-3GalNAc, mediator of the anti-proliferative effects of the protein,
binds at a shallow depression on the surface of the molecule. The lectin
has two distinct binding sites per monomer that recognize the different
configuration of a single epimeric hydroxyl (Carrizo et al. 2005).

Aqueous extracts of the sporophores of eight mushroom species have

been assessed for their ability to prevent H2O2-induced oxidative damage
to cellular DNA using the single-cell gel electrophoresis ("Comet") assay.
The highest genoprotective effects were obtained with cold (20ºC) and
hot (100ºC) water extracts of Agaricus bisporus and Ganoderma lucidum
fruit bodies, respectively. These edible mushrooms therefore represent a
valuable source of biologically active compounds with potential for
protecting cellular DNA from oxidative damage (Rocha et al. 2002). A
heat-labile protein has also been identified in fruit bodies of Agaricus
bisporus which protects Raji cells (a human lymphoma cell line) against
H2O2-induced oxidative damage to cellular DNA (Shi et al. 2002).

Lectins from Agaricus bisporus and Agaricus campestris have been shown
to stimulate insulin and glucagon release from isolated rat islets in the
presence of 2 mM glucose. Maximal stimulation of insulin release was
reported at lectin concentrations above 58mg/mL (approximately 1mM).
The lectin did not alter islet glucose oxidation to CO2 or incorporation of
[3H] leucine into trichloracetic acid-precipitable material, nor did it modify
rates of insulin secretion induced by 20 mM glucose. None of nine other
lectins tested stimulated insulin release, whereas stimulation of fat cell
glucose oxidation was a general property of the lectins. The data also
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suggesting that lectin binding is essential for the expression of insulin-
releasing activity. The authors proposed that the specific interaction
between mushroom lectin and its receptors may lead to conformational
changes in the structure of the membranes of the islet A2- and B-cells
that facilitate exocytosis (Ewart et al. 1975).

Agaricus bisporus and Pleurotus sajor caju have been assayed in vitro for
their anti-microbial activities using aqueous and organic solvents
extracts. It has been shown that Escherichia coli 390, Escherichia coli
739, Enterobacter aerogenes, Pseudomonas aeruginosa and Klebsiella
pneumoniae were most sensitive to aqueous, ethanol, methanol and
xylene extracts of these mushrooms (Tambekar et al. 2006).

A report on the fractionation of extracts of the edible mushroom,

Volvariella volvacea, has shown the isolation of two heterocyclic
carboxylic acids, namely pyridine-3-carboxylic acid [nicotinic acid] and
pyrazole-3(5)-carboxylic acid and four steroidal metabolites ergosterol,
5-dihydroergosterol, ergosterol peroxide, and cerevisterol. In light of the
structural similarity of pyrazole-3(5)-carboxylic acid to pyrazole-3-
carboxylic acids, which act as agonists for nicotinic acid receptors, the
levels of pyridine-3-carboxylic acid and pyrazole-3(5)-carboxylic acid
were estimated in V. volvacea and two other edible mushrooms, namely
Agaricus bisporus and Calocybe indica. Significant levels of pyridine-3-
carboxylic acid (nicotinic acid) were found in C. indica, and pyrazole-3(5)-
carboxylic acid was found in abundance in A. bisporus. Correlations have
been suggested between the occurrence of these compounds in
mushrooms and consumption as well as beneficial health effects
(Mallavadhani et al. 2006).

Plasma cholesterol concentration in rats has been shown to be reduced

by feeding of mushroom (Agaricus bisporus) fiber. The results
demonstrated that mushroom fiber (and sugar beet fiber) lowered the
serum total cholesterol level by enhancement of the hepatic low density
lipoprotein (LDL) receptor mRNA (Fukushima et al. 2000). Similar
cholesterol-lowering effects in rats of Maitake (Grifola frondosa) fiber,
Shiitake (Lentinus edodes) fiber, and Enokitake (Flammulina velutipes)
fiber have also been reported (Fukushima et al. 2001).

Long term Agaricus bisporus consumption has been studied in rats.

Female Charles River Sprague - Dawley rats were fed a diet containing a
30% dry powder of A. bisporus for 500 days. A control group was given a
basal diet without A. bisporus. There was no significant difference in the
incidence of tumours between the experimental group and control group.
No carcinogenic activity of A. bisporus was observed in this long-term
study (Matsumoto et al. 1991).
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A study by Toth and co-workers in which Agaricus bisporus, baked at
220-230ºC for 10 minutes and subsequently fed to mice for 12h each
day, five days each week throughout their life and also fed a well-
balanced semi-synthetic diet for 12h each day for five days and for the
remaining two full days each week, showed no statistically significant
difference in tumours in the lungs, blood vessels, cecum, and colon when
compared to the untreated controls. The estimated average daily
mushroom consumption per animal was 4.8g for female mice and 4.2g
for male mice (Toth et al. 1997), which exceeds the average daily
consumption of mushrooms by humans (Shephard et al. 1995).

Lectins from Agaricus bisporus, Phaseolus vulgaris, Momordica charantia,

Ricinus communis and its constituent chains have been shown to inhibit
human immunodeficiency virus type 1 (HIV-1) reverse transcriptase
(Wang and Ng, 2001), while phenolic compounds present in extracts from
A. bisporus strongly generated reactive oxygen species (suggesting
immunomodulatory effects) in human PBMCs and K 562 cells in vitro(Wei
et al. 2008).

Agaricus blazei
In general, the anti-tumour activity of
Agaricus blazei appears to be mainly due to
the activation of the immune system rather
than to any direct effects on tumour cells.
This is supported by the fact that
macrophages derived from rat bone marrow
have been shown to be activated and
cytokines such as tumour necrosis factor-
alpha (TNF-a), interleukin-1 (IL-1) and IL-8,
and nitric oxide (NO) were secreted, in
response to water extracts in in vitro experiments. Furthermore, oral
administration of Agaricus blazei water extracts to mice has been shown
to induce the activation of macrophages and T cells in vivo. Anti-
genotoxic, anti-mutagenic and anti-clastogenic effects have also been
detected in Agaricus blazei water extracts (Sorimachi and Koge, 2008).
Agaricus blazei Murrill extracts, under certain conditions, have also been
shown to have anti-mutagenic activities in mice that may contribute to an
anti-carcinogenic effect (Delmanto et al. 2001).
Oral administration of dried fruiting bodies of A. blazei has been shown to
augment cytotoxicity of natural killer (NK) cells in wild-type (WT)
C57BL/6, C3H/HeJ, and BALB/c mice. Augmented cytotoxicity was
demonstrated by purified NK cells from treated wild-type (WT) and RAG-
2-deficient mice, but not from interferon-gamma (IFN-gamma) deficient
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mice. NK cell activation and IFN-gamma production was also observed in
vitro when dendritic cell (DC)-rich splenocytes of WT mice were
coincubated with an extract of A. blazei. Both parameters were largely
inhibited by neutralizing anti-interleukin-12 (IL-12) monoclonal antibody
(mAb) and completely inhibited when anti-IL-12 mAb and anti-IL-18 mAb
were used in combination. An aqueous extract of the hemicellulase-
digested compound of A. blazei particle (ABPC) induced IFN-gamma
production more effectively, and this was completely inhibited by anti-IL-
12 mAb alone. NK cell cytotoxicty was augmented with the same
extracts, again in an IL-12 and IFN-gamma-dependent manner. These
results demonstrate that A. blazei and ABPC augmented NK cell
activation through IL-12-mediated IFN-gamma production (Yuminamochi
et al. 2007).

Aqueous extracts of Agaricus blazei fruiting body prepared at different

temperatures have been fractionated by ethanol precipitation with
various ethanol concentrations. The original aqueous extracts of A. blazei
failed to stimulate natural killer (NK) cell activity in murine spleen cells in
vitro, but the strongest effect was observed in a 30% ethanol-soluble-
50% ethanol-insoluble fraction prepared from the extract at 40ºC
(fraction A-50). Fraction A-50 also showed the strongest augmenting
effect on interferon (IFN)-gamma production. This augmentation of NK
activity and IFN-gamma production by fraction A-50 was completely
abrogated by heat treatment (Zhong et al. 2005).

Clinical effects and safety evaluation of Agaricus Blazei Condensed Liquid

(Agaricus Mushroom Extract; ABCL) on human volunteers with C-type
hepatitis has been studied. A total of 20 patients (10 male, 10 female)
with chronic C-type hepatitis received the ABCL orally twice per day for 8
weeks. No toxicological effects, nor other side effects were observed
(Inuzuka and Yoshida, 2002).

The effects of protein-bound polysaccharides (A-PBP and L-PBP),

extracted from the mycelia of Agaricus blazei and Lentinus edodes, on
serum cholesterol and body weight have been investigated in 90 female
volunteers. The data demonstrated a weight-controlling and
hypolipidemic effect of both A-PBP and L-PBP via a mechanism involving
absorption of cholesterol (Kweon et al. 2002).

Polysaccharide fractions of Agaricus blazei have been prepared from

cultured A. blazei by repeated extraction with hot water (AgHWE), cold
NaOH (AgCA), and then hot NaOH (AgHA). By chemical, enzymic, and
NMR analyses, the primary structures of AgHWE, AgCA, and AgHA were
mainly composed of 1,6-beta-glucan. Among these fractions, the NaOH
extracts showed anti-tumour activity against the solid form of Sarcoma
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180 in ICR mice. To demonstrate the active component in these fractions,
several chemical and enzymic treatments were applied. These fractions
were found to be i) neutral beta-glucan passing DEAE-Sephadex A-25, ii)
resistant to periodate oxidation (I/B) and subsequent partial acid
hydrolysis (I/B/H), iii) resistant to a 1,3-beta-glucanase, zymolyase,
before I/B, but sensitive after I/B/H. In addition, after I/B/H treatment of
the neutral fraction of AgCAE, a signal around 86 ppm attributable to 1,3-
beta glucosidic linkage was detectable in the 13C-NMR spectrum. These
data strongly suggest that a highly branched 1,3-beta-glucan segment
forms the active centre of the anti-tumour activity (Ohno et al. 2001).

Agaricus blazei Murill has been reported to possess biological effects that
include immunomodulatory activities, although the number of in vivo
studies is limited. A recent study has evaluated the immunomodulatory
effects of A. blazei in 160 male Balb/cByJ mice. The mice were divided
into four groups and treated with various quantities of intragastric A.
blazei extract or distilled water for 8 to 10 weeks. Nine parameters,
relating to general immune function or adaptive immunity against
immunogen chicken ovalbumin, were determined. The mice receiving A.
blazei extract exhibited significantly greater serum immunoglobulin G
levels, increased T-cell numbers in spleen, and elevated phagocytic
capability compared with controls. Consumption of A. blazei was also
associated with significant increases in ovalbumin-specific serum
immunoglobulin G level, delayed-type hypersensitivity, splenocyte
proliferation rate, and tumour necrosis factor-alpha secretion by
splenocytes, indicating that A. blazei Murill possesses a wide range of
immunomodulatory effects in vivo (Chan et al. 2007). Agaricus blazei
has also been reported to have inhibitory effects on mast cell-mediated
anaphylaxis-like reactions (Choi et al. 2006b).

Beta-glucans and and their enzymatically hydrolyzed oligosaccharides

from Agaricus blazei have anti-hyperglycemic, anti-hypertriglyceridemic,
anti-hypercholesterolemic, and anti-arteriosclerotic activity indicating
overall anti-diabetic activity in diabetic rats. However, the enzymatically
hydrolyzed oligosaccharides have been shown to have around twice the
activity of beta-glucans with respect to anti-diabetogenic activity (Kim et
al. 2005).

Extracts from Agaricus blazei Murill (AbM) have been evaluated on

changes to gene expression on a human monocyte cell line (THP-1).
Changes in the levels of mRNA transcripts were measured using 35 k
microarrays, and the changes in select cytokine gene products by
immunoassays. Lipopolysaccharide (LPS) was included for comparison.
Both AbM and LPS had very significant effects on gene expression. Genes

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related to immune function were selectively up-regulated, particularly
pro-inflammatory genes such as the interleukins IL1B and IL8. Although
most genes induced by AbM were also induced by LPS, AbM produced a
unique profile, e.g., as to a particular increase in mRNA for the cytokines
IL1A, CXCL1, CXCL2 and CXCL3, as well as PTGS2 (cyclooxygenase2)
(Ellertsen et al. 2006)

An extract from Agaricus blazei Murill Kyowa (ABMK), has been reported
to possess anti-mutagenic and anti-tumour effects. A study has
investigated the effects of ABMK consumption on immunological status
and quality of life in cancer patients undergoing chemotherapy. One
hundred cervical, ovarian, and endometrial cancer patients were treated
either with carboplatin (300mg/m2) plus VP16 (etoposide, 100mg/m2) or
with carboplatin (300mg/m2) plus taxol (175mg/m2) every 3 weeks for at
least three cycles, with or without oral consumption of ABMK. The
authors observed that natural killer cell activity was significantly higher in
the ABMK-treated group compared to the non-treated placebo group (n =
61). However, no significant difference in lymphokine-activated killer and
monocyte activities was observed in a manner similar to the count of
specific immune cell populations between ABMK-treated and non-treated
groups. However, chemotherapy-associated side effects such as appetite,
alopecia, emotional stability, and general weakness were all reported to
be improved by ABMK treatment, with the authors suggesting that ABMK
treatment may have some beneficial effects for gynecological cancer
patients undergoing chemotherapy (Ahn et al. 2004).

Sodium pyroglutamate isolated from Agaricus blazei has been shown to

have potent anti-tumour and anti-metastatic actions, as well as immune-
modulatory activity, in tumour-bearing mice (Kimura et al. 2004).

Oral administration of ergosterol, isolated from the lipid fraction of

Agaricus blazei Murill has also been shown to have anti-tumour activity in
Sarcoma 180-bearing mice. Ergosterol reduced tumour growth at doses
of 400 and 800mg/kg. Administration of ergosterol for 20 days was
reported to be without side effects, such as decreases in body,
epididymal adipose tissue, thymus, and spleen weights and leukocyte
numbers induced by cancer chemotherapy drugs. Ergosterol had no
cytotoxicity against tumour cells and it appears as though the antitumour
activity of ergosterol might be due to direct inhibition of angiogenesis
induced by solid tumours (Takaku et al. 2001).

The effects of low molecular weight products extracted from Agaricus

blazei Murill on MethA tumour cell growth have been studied. Inoculation
of a low molecule fraction (LM) into the primary tumour of a two-tumour
mouse model resulted in a marked inhibition of the tumour, not only in
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the right flank, but also in the non-injected left flank. Chromatographic
purification and physicochemical characterization showed the main
tumouricidal activity to be located in a low molecule fraction-3 (LM-3),
containing alpha-1,4-glucan-beta-1,6-glucan complex with an average
molecular weight of 20kDa. Serum levels of immunosuppressive acidic
protein (IAP) in mice receiving LM fractions, particularly LM-3,
significantly increased, indicating the possible activation of granulocytes
(Fujimiya et al. 1999).

Induction of apoptosis in human gastric epithelial AGS cancer cells by an

aqueous extract of Agaricus blazei has been demonstrated. It was found
that an Agaricus blazei extract could inhibit cell growth in a dose-
dependent manner, which was associated with the arrest of G2/M phase
and the induction of apoptotic cell death via caspase-3 activation (Jin et
al. 2006). A subsequent study by the same group has shown a similar
effect on apoptosis by Agaricus blazei on human leukemic U937 cells via
similar mechanisms (regulation of Bcl-2 and caspase-3) (Jin et al.
2007).

The effect of an RNA-protein complex isolated from A. blazei Murill, on

human leukemia HL-60 cells has been studied. Typical apoptotic
characteristics were determined by morphological methods using DNA
agarose gel electrophoresis and flow cytometry. The data showed that
the fraction from Agaricus blazei induced HL-60 cell apoptosis and that
the combined effect of down-regulation of telomerase activity and up-
regulation of mRNA expression of the caspase-3 gene could be the
primary mechanism of induction of apoptosis. These findings provide
good evidence that the isolated fraction may be of value for the clinical
treatment of acute leukemia (Gao et al. 2007).

An extract from fruit bodies of Agaricus blazei has been evaluated in a

"double grafted tumour system" mouse model and reported to cure the
primary tumour and inhibit the growth of metastatic tumours in this
model. A separate extract from Agaricus blazei (Himematsutake)
inhibited the growth of the primary tumour. An immuno-suppresive acidic
protein (IAP) was induced by both the Agaricus and Himematsutake
preparations but not by Lentinus edodes. Lentinus edodes had no effect
on the growth of either the primary or metastatic tumours (Ebina, 2005).

An aqueous extract of Agaricus blazei has been shown to exert a hepato-

protective effect on both liver toxicity and hepato-carcinogenesis on rat
liver toxicity induced by different doses of diethylnitrosamine (Barbisan et
al. 2002).

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A subsequent study from another group has also demonstrated the
chemo-preventative potential of an Agaricus blazei (Ab) Murrill
mushroom meal in a medium-term rat liver carcinogenesis assay. Male
Wistar rats initiated for hepatocarcinogenesis with diethylnitrosamine
(DEN, 200mg/kg i.p.) were fed during a 6-week period with dry
powdered mushroom strains Ab 29 or 26, each one with opened (OB) or
closed basidiocarp (CB), mixed at a level of 10% in a basal diet. Chemo-
preventative activity of the mushroom meal was observed for the Ab 29
(OB and CB) and Ab 26 (CB) strains in terms of the number of putative
pre-neoplastic altered foci of hepatocytes which express either the
enzyme glutathione S-transferase, placental form (GST-P+) or the
transforming growth factor-alpha, and for the Ab 29 (OB) and Ab 26 (CB)
strains on the size of GST-P+ foci. This was associated with inhibition of
foci cell proliferation in the animals fed the Ab 29 (OB) and Ab 26 (CB)
strains. The results suggest that the protective influence of the Ab meal
against the DEN potential for rat liver carcinogenicity depends on both
the strain and period of mushroom harvest (Pinheiro et al. 2003).

The effects of crude extracts of the mushroom Agaricus blazei Murrill

(Agaricaceae) on both DNA damage and placental form glutathione S-
transferase (GST-P)-positive liver foci induced by diethylnitrosamine
(DEN) have also been investigated in adult male Wistar rats. The data
indicated that previous treatment with the highest concentration of
Agaricus blazei (11.5mg/ml) significantly reduced DNA damage,
indicating a protective effect against DEN-induced liver
cytotoxicity/genotoxicity (Barbisan et al. 2003a) while in a subsequent
study, the same group reported that treatment with aqueous extracts of
Agaricus blazei does not exert a protective effect against the
development of GST-P-positive foci induced by DEN (Barbisan et al.
2003b).

Anti-bacterial effects of Agaricus blazei Murill (AbM) have been

investigated. The AbM extract protected against systemic Streptococcus
pneumoniae 6B infection in mice and was most effective when given 24h
before inoculation but it also had protective effects when given together
with challenge compared with control. The lack of an antibiotic effect on
pneumococci in vitro and increased levels of cytokines MIP-2 and TNF in
the serum of mice receiving AbM extract, indicated that the protective
effect of AbM was due to the involvement of the native immune system.
The anti-infection properties of AbM have therefore been shown in vivo
and the results suggest that AbM extract may be useful as an additional
prophylactic and possibly therapeutic treatment against bacterial
infections in humans (Bernardshaw et al. 2005).

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A subsequent study by the same group has shown that an extract of
Agaricus blazei Murill can protect against lethal septicemia in a mouse
model of faecal peritonitis. Bacterial septicemia can occur during
gastroenterological surgery. The putatively anti-infective
immunomodulatory action of Agaricus blazei Murill (AbM) has been
studied in an experimental peritonitis model in BALB/c mice. The mice
were orally given an extract of AbM or phosphate-buffered saline 1 day
before the induction of peritonitis with various concentrations of faeces
from the mice. The state of septicemia, as measured by the number of
colony-forming units of bacteria in blood, and the survival rate of the
animals were compared between the groups. Mice that were orally
treated with Agaricus blazei Murill extract before bacterial challenge
showed significantly lower levels of septicemia and improved survival
rates (Bernardshaw et al. 2006).

A randomized, double-blinded, and placebo-controlled clinical trial has

evaluated the effects of Agaricus blazei Murill in combination with
metformin and gliclazide on insulin resistance in type 2 diabetes.
Supplementation of Agaricus blazei Murill extract improved insulin
resistance among subjects with type 2 diabetes. The increase in
adiponectin concentration after taking Agaricus blazei Murill extract for 12
weeks may be the mechanism that results in the observed effect (Hsu et
al. 2007).

A recent study has demonstrated that ultrafiltration, in combination with

spray-drying, is applicable for the preparation of protein-bound
polysaccharide powders with higher anti-tumour activities from Agaricus
Blazei Murill (Hong et al. 2007). Thermostable antioxidant activity has
also been reported from Agaricus blazei Murill (Izawa and Inoue, 2004).

A study to evaluate the chronic toxicity and oncogenicity of Agaricus

blazei Murill in F344 rats has been reported (Lee et al. 2008). Long-
term (2 years) feeding of rats of a powdered diet containing Agaricus
blazei at levels up to 25,000 ppm (parts per million) revealed no
remarkable change in mean body weight, body weight gain, hematologic
or serum chemistry parameters, or absolute or relative organ weights in
control or treatment groups. Mortality in male treatment (mushroom)
groups was significantly lower than in controls. Histopathological studies
showed no increased incidence of tumours.

A recent study has also reported good bioavailablity of both copper and
zinc from mycelium of Agaricus blazei Murrill equating to very good levels
of recommended daily intakes of these minerals from small amounts of
(1g) of this mushroom (Rabinovich et al. 2007).

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Anti-viral activities of Agaricus blazei Murill have been demonstrated
against cytopathic effects induced by western equine encephalitis (WEE)
virus by the mycelial fractions but not those of fruiting bodies (Sorimachi
et al. 2001).

Ethanol extracts and hot water extracts of Agaricus blazei, Agrocybe

cylindracea, and Boletus edulis have been shown to have significant
antioxidant properties. The ethanolic extracts were more effective than
hot water extracts in antioxidant activity using the conjugated diene
method and scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals
whereas hot water extracts were more effective in reducing power,
scavenging ability on hydroxyl radials and chelating ability on ferrous ions
as demonstrated by their lower EC50 values. Naturally occurring
antioxidant components including total tocopherols (3.18-6.18mg/g) and
total phenols (5.67-5.81mg/g) were found in the extracts and their
contents were associated with the EC50 value of the antioxidant
properties (Tsai et al. 2007).

Differences of the pharmacological effects of Agaricus blazei cultured on

various materials have been examined. Agaricus blazei mushrooms were
prepared on culture media composed of 1) tops of sugar cane shoots
(stems and leaves), 2) rice straw 3) wheat straw, 4) broad leaf tree bark,
and 5) used substrate after Pleurotus ostreatus cultivation. The
pharmacological effects of this mushroom were examined by the
following methods; 1) anti platelet aggregation stimulated by PAF or
arachidonic acid Na, 2) inhibition of IL-8 gene expression stimulated by
TNF-alpha, 3) improvements of rough surfaces by using replica method.
In both the anti-platelet aggregation test and chemokine gene revelation
control test, A. blazei cultured on the top shoot of sugar cane medium
showed the most effective results compared with that cultured on other
media. The results suggested that the A. blazei cultured on the top shoot
of sugar cane medium has increased pharmacological activity compared
to mushrooms cultured on rice or wheat straw or broad leaf bark
(Yoshimoto et al. 2005).

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Agaricus brasiliensis
Agaricus brasiliensis (previously named Agaricus
blazei ss. Heinem), also known as the sun
mushroom, is native to Southeastern Brazil, and
is widely consumed, mainly in the form of tea.
Aqueous (AqE) and ethanol (EtOHE) extracts and
an isolated polysaccharide (PLS) from the fruiting
body of A. brasiliensis have been evaluated for
anti-viral activity against poliovirus type 1 in
HEp-2 cells. The evaluation of the time of
addition by plaque assay showed that when AqE,
PLS and EtOHE were added, just after the virus
inoculation (time 0 h), there was a
concentration-dependent reduction in the
number of plaques by up to 50%, 67% and 88%,
respectively. The test substances showed anti-viral activity and were
more effective when added during the poliovirus infection. As the extracts
had little effect on reducing viral adsorption and did not show any
virucidal effect, the authors suggested that they may act at the initial
stage of the replication of poliovirus (Faccin et al. 2007).

The mycelium polysaccharide and exo-polysaccharide (EPS) of Agaricus

brasiliensis LPB 03 produced by submerged fermentation has shown
strong inhibition against Sarcoma 180 in mice, reaching 72.19%
inhibition compared to a control group. Furthermore, 50% of mice in the
test group demonstrated total tumour regression (Fan et al. 2007).

The structure and anti-tumour activity of polysaccharide fractions of the

fruit body of Agaricus brasiliensis have been studied in cold and hot water
extracts (CWE and HWE) on a mouse diabetic model (C57BL Ksj-db/db).
Compared to the water administered control group, the body weight,
urinary glucose exclusion, urinary pH, blood glucose level, and organs
weight were comparable. The splenocytes of CWE administered mice
produced a higher concentration of interleukin-6. By megascopic and
microscopic examinations of renal sections, the number of the mice
having abnormal kidney was 3/5 (control), 2/5 (HWE), and 0/5 (CWE)
suggested the activity of the renal protection in the cold water extract.
The results strongly suggest that the pharmacological action of the cold
water extract of A. brasiliensis is significantly stronger than that of the
hot water extract (Furukawa et al. 2006).

The administration of ethanol-soluble, boiling water-soluble, and

ammonium oxalate-insoluble fractions isolated from the fruiting bodies of
13
Agaricus brasiliensis into C3H/HeN mice has been shown to play an
important role in the maintenance of hematopoietic cells for compromised
patients at the risk of infection associated with malignancy (Fujimiya et
al. 2006).

Phenolic compounds present in mushroom extracts from A. brasiliensis

strongly generated reactive oxygen species (suggesting
immunomodulatory effects) in human PBMCs and K 562 cells in vitro (Wei
et al. 2008).

Agrocybe aegerita (Black Poplar, Piopinno, Chiodini)

Agrocybe aegerita is an edible

mushroom with reported anti-tumour
properties. An investigation of
bioactivity gave positive results for
ceramide, methyl-beta-D-
glucopyranoside and alpha-D-
glucopyranoside, along with linoleic
acid and its methyl ester. Ceramide
inhibited the cyclooxygenase (COX)
enzymes, COX-1 and -2 and its anti-
cancer potential was investigated against five human cancer cell lines in
vitro and it was found to inhibit the proliferation of stomach, breast and
central nervous system cancer cell lines suggesting that the compound
may be useful in alleviating inflammatory conditions, as well as possibly
reducing the development of the above cancers (Diyabalanage et al.
2008).

A lectin from Agrocybe aegerita (AAL) has been found to possess potent
tumour-suppressing function and tumour cell apoptosis-inducing activity.
Its full sequence has been published. It has been reported that AAL is a
member of the galectin family and the dimeric form is the active unit for
functional performance. The recombinant AAL showed comparable
tumour cell apoptosis-inducing activity with the wild AAL but no DNase
activity (Yang et al. 2005).

Antioxidant activity of a methanol crude extract and its fractions, from

the fruiting bodies of Agrocybe aegerita, has been evaluated by
scavenging activity of 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic
acid) radical cation (ABTS(+)) and inhibition of lipid peroxidation of rat
brain homogenate. The ethyl acetate (EA) fraction, which showed the
most potent antioxidant activity in the above two assays, was further

14
fractionated by a Sephadex LH-20 column into four subfractions (EA1-
EA4). Significant correlation was found between the total phenolic
content and the antioxidant activity in the EA fraction and its sub-
fractions (Lo and Cheung, 2005).

Proliferation of human leukemic U937 cells has been shown to be

significantly inhibited by conditioned medium of human peripheral blood
mononuclear cells stimulated with cold-water extracts (10-800 mg/mL
of medium) of dietary mushrooms, Hypsizigus mamoreus, Agrocybe
aegerita and Flammulina velutipes (Ou et al. 2005).

Agrocybe chaxingu
Osteoclast forming suppressive compounds
(important in osteoporosis) have been
isolated from the mushroom Agrocybe
chaxingu (Abel et al. 2007).

Auricularia auricular (Wood Ear)

In vitro evaluation of antioxidant activities of
Auricularia auricular has shown significant inhibition
of lipid peroxidation, and potent hydroxyl radical
scavenging activity when compared with the drug
catechin. The IC50 value of crude, boiled and
ethanolic extracts of A. auricula represented 403,
510, and 373mg/ml respectively of hydroxyl radical
scavenging activity and 310, 572 and 398mg/ml
respectively of lipid peroxidation, while crude,
boiled and ethanolic extracts were shown to
significantly increase nitric oxide production (664,
191 and 850pmole/mg dry wt/h respectively) over the control (Acharya
et al. 2004).

Auricularia auricular has been shown to have hypocholesterolemic

properties (Cheung, 1996b).

Constipation is one of the most prevalent gastrointestinal complaints and

high fiber intake is recommended as an initial therapy for constipation.
Ear mushrooms (Auricularia) are known to have higher fiber contents (by

15
~50%) than other mushroom varieties. In patients with functional
constipation, fiber supplements using ear mushrooms has been shown to
significantly improve constipation related symptoms without serious side
effects (Kim et al. 2004).

An acidic polysaccharide with anti-coagulant activity has been isolated

from Auricularia auricula using water, alkali or acid extracts. The alkali
extract showed the highest anti-coagulant activity and was further
purified using gel filtration chromatography. The specific anti-coagulant
activity of the purified polysaccharide was 2 international units (IU)/mg
and its average mass was approximately 160kDa. The polysaccharide
from this species of mushroom contains mainly mannose, glucose,
glucuronic acid and xylose but no sulfate esters. Its anti-coagulant
activity was due to catalysis of thrombin inhibition by anti-thrombin, but
not by heparin cofactor II. Inhibition of Factor Xa by anti-thrombin was
not catalyzed by the polysaccharide. The glucuronic acid residues were
essential for the anti-coagulant action of the mushroom polysaccharide
since the activity disappeared after reduction of its carboxyl groups. In ex
vivo tests using rats orally fed with the polysaccharide, an inhibitory
effect on platelet aggregation was observed, as with aspirin, a well-
known anti-platelet agent. The authors suggested that polysaccharides
from these mushrooms may constitute a new source of compounds with
action on coagulation, platelet aggregation and, perhaps, on thrombosis
(Yoon et al. 2003) .

Auricularia polytricha (Jew's Ear)

An immunomodulatory protein (APP) has been
purified from the fruiting body of an edible Jew's
Ear mushroom, Auricularia polytricha, by
extraction using 5% cold acetic acid in the
presence of 0.1% 2-mercaptoethanol, followed
by ammonium sulfate fractionation, DE-52 and
P,MonoQ anion-exchange chromatography. The
molecular mass of APP was around 13.4kDa and
its pI is approximately 5.1. APP is a simple
protein without carbohydrate, and can
agglutinate mouse red blood cells. APP alone activates murine
splenocytes, markedly increasing their proliferation and gamma-
interferon (IFN-gamma) secretion, and presented no cytotoxicity in vitro.
Although murine splenocytes are stimulated by the mitogen concanavalin
A (ConA), APP suppressed their proliferation in a dose-dependent
manner. APP also enhanced the production of both nitric oxide (NO) and
tumour necrosis factor-alpha (TNF-alpha) by LPS-induced RAW 264.7
16
macrophages. The data suggest that this protein isolated from Auricularia
polytricha is an immune stimulant (Sheu et al. 2004).

Cordyceps sinensis (Caterpillar mushroom)

An exo-polysaccharide fraction from cultivated
Cordyceps sinensis has been shown to have
immunomodulatory and anti-tumour effects on (B16
melanoma) tumour-bearing mice (Zhang et al. 2005).

Cultivated mycelium of Cordyceps sinensis, sequentially

extracted by petroleum ether (PE), ethyl acetate
(EtOAc), and ethanol (EtOH) showed a significant and
dose-dependent inhibitory effect on the proliferation of
four cancer cell lines, MCF-7 breast cancer, B16 mouse
melanoma, HL-60 human pre-myelocytic leukemia and
HepG2 human hepatocellular carcinoma, with IC50
values below 132 mg/ml. A hot water extract failed to
show such activity. The EtOAc extract, in particular, had the most potent
effect against all four cancer cell lines, with IC50 values between 12mg/ml
(on B16) and 45mg/ml (on MCF-7). In contrast, it had much lower
cytotoxicity against normal mouse bone marrow cells. The EtOAc extract
contained carbohydrates, adenosine, ergosterol and a trace amount of
cordycepin, of which ergosterol and related compounds were identified as
a major class of active constituents contributing to the in vitro
cytotoxicity. In an animal test, the EtOAc extract showed a significant
inhibiting effect on B16-induced melanoma in C57BL/6 mice, causing a
60% decrease of tumour size over 27 days (Wu et al. 2007).

An extract of Cordyceps sinensis (CSE) has been tested in C57BL/6 mice

implanted subcutaneously with syngeneic EL-4 lymphoma cells. Oral
administration of the extract led to a reduction of tumour size and
prolongation of the host survival time. As for the activities of peritoneal
macrophages, chemotaxis was dramatically depressed within a few days
after EL-4 transplantation up to the end of life, but treatment with CSE at
-14, -7, -4, +4, +7 and +10 days after the tumour transplantation
augmented the activity about four times greater than that of the control.
Phagocytic activity of macrophages was also decreased in tumour-bearing
mice treated with cyclophosphamide (100mg/kg) 3 and 5 days after
tumour transplantation, but administration of CSE restored the activity to
more than the normal level. The overall efficacy of CSE was tested with
protective activity against systemic infection by Salmonella enteritides.
The tumour-bearing mice receiving this extract lived significantly longer
than the other groups without CSE (Yamaguchi et al. 1990).

17
A study has been conducted to investigate a hypocholesterolemic effect
of a hot-water fraction (HW) from cultured mycelia of Cordyceps sinensis.
In mice fed a cholesterol-free diet and those fed a cholesterol-enriched
diet, body and liver weights were not significantly different from those of
the controls. The serum total cholesterol (TC) of all mice groups
administered HW (150 and 300mg/kg/d, respectively) with the
cholesterol-enriched diet decreased more than in the control group.
Among the mice fed the cholesterol-enriched diet, HW also increased the
high-density lipoprotein (HDL) cholesterol level, but decreased the very
low-density lipoprotein plus low-density lipoprotein (VLDL+LDL)
cholesterol level. The changes in HDL-and VLDL+LDL-cholesterol levels
consequently decreased the atherogenic value. The results indicated that
HW in rats administered a cholesterol-enriched diet decreased the plasma
cholesterol level. The 300mg/kg dose had a significant effect on the
serum total cholesterol level (Koh et al. 2003).

Flammulina velutipes (Enoki)

It has been reported that there is

currently no effective therapy for
malignant estrogen-independent breast
cancer. Screening of 38 species of edible
mushrooms on human estrogen-receptor
positive (ER+) (MCF-7) and estrogen-
receptor negative (ER-) (MDA-MB-231,
BT-20) breast cancer cells has been
undertaken to select potential agents with
broad-spectrum anti-tumour activity
against breast cancer cells. Water-based
extracts of three mushroom species, Coprinellus sp., Coprinus comatus
and Flammulina velutipes (CME, CCE and FVE, respectively), have been
identified as anti-breast cancer agents. The anti-tumour activities
included marked growth inhibition of both ER+ and ER- breast cancer
cells, induction of rapid apoptosis on both ER+ and ER- cells, and
significant inhibition of MCF-7 tumour colony formation in vitro. The anti-
proliferative and cytotoxic activities of the three mushroom extracts were
dose-dependent, regardless of the hormone receptor status of the cancer
cells. The degree of produced cytotoxicity on ER- breast cancer cells was
very high. Mushroom extracts CME and FVE induced a rapid (within 5
hours) apoptosis on MCF-7 and MDA-MB-231 cells. MCF-7 tumour colony
formation rate was reduced by 60% in CCE- and CME-treated cells and
nearly completely inhibited (99%) by FVE treatment. These results
suggest that the mushroom species Coprinus comatus, Coprinellus sp.
18
and Flammulina velutipes contain potent anti-tumour compounds for
breast cancer. This finding is important due to the lack of
chemotherapeutic and chemo-preventive agents for ER- human breast
cancer (Gu and Leonard, 2006).

An anti-tumour polysaccharide, EA3 isolated from Flammulina velutipes

(CURT. ex FR.) SING. has been shown to be composed of D-glucose with
the chemical structure of a beta-(1 leads to 3)-glucan. Another anti-
tumour polysaccharide (EA5) also isolated from F. velutipes was
fractionated and among the polysaccharides isolated, the highest
molecular weight polysaccharide (EA501) showed the highest anti-tumour
activity. The component sugars of EA501 were found to be D-glucose
42.3%, D-galactose 17.3%, D-mannose 12.2%, D-xylose 6.7% and L-
arabinose 14.7% (Ikekawa et al. 1982).

A further study by the same group has shown that proflamin, isolated
from the culture mycelium of Flammulina velutipes (Curt. ex Fr.) Sing. is
a weakly acidic glycoprotein containing more than 90% protein and less
than 10% carbohydrate, and has a molecular weight of ~13,000Da.
Proflamin has been shown to be markedly effective against the syngeneic
tumours, B-16 melanoma (B-16) and adenocarcinoma 755 (Ca-755) in
the mouse. The increases in median survival time of treated mice with B-
16 and Ca-755 were 86% and 84%, respectively. Proflamin exhibited no
cytocidal effect against the cultured cell lines in vitro. Oral administration
of proflamin produced no lethal or any other apparent adverse effect in
mice (Ikekawa et al. 1985).

Antioxidant activity of submerged cultured mycelium extracts of higher

Basidiomycetes mushrooms has recently been reported. Antioxidant
properties were studied from 28 submerged cultivated mycelium
Basidiomycetes strains of 25 species. Three solvents - ethanol, water
(culture liquid), and ethyl acetate were used for extraction. Water
extracts from Coprinus comatus, Agaricus nevoi, and Flammulina
velutipes (Enoki) showed high antioxidant activities (AA) at 2mg/ml.
When the ethanol extracts were tested, the highest AA were found in
Agaricus nevoi, Omphalotus olearius, and Auricularia auricula-judae
extracts at a concentration of 2mg/ml. The AA of ethanol extracts from
Agrocybe aegerita and C. comatus increased from 46.6% to 82.7% and
from 2.4% to 62.1%, respectively, when the concentration of the extract
increased from 2mg/ml to 4-8mg/ml with the authors suggesting that the
extracts could be suitable as antioxidative agents and bioproducts
(Asatiani et al. 2007a).

Plasma cholesterol concentration in rats has been shown to be reduced

by feeding of mushroom Enokitake (Flammulina velutipes) fiber. The
19
results demonstrated that mushroom fiber lowered the serum total
cholesterol level by enhancement of the hepatic low density lipoprotein
(LDL) receptor mRNA (Fukushima et al. 2001).

Angiotensin-converting enzyme (ACE) inhibitory activity (which has an

effect on blood pressure reduction) has been demonstrated in the culture
broth from Flammulina velutipes (strain 414). Nutritional requirements
for the production of ACE inhibitory activity from F. velutipes were shown
to include sucrose, ammonium acetate, and glutamic acid (Kim et al.
2002).

Two cuparene-type sesquiterpenes, enokipodins C (1) and D (2), have

been isolated from culture medium of Flammulina velutipes (Enoki),
along with enokipodins A (3) and B (4). All the metabolites showed anti-
microbial activity against the fungus Cladosporium herbarum, and gram-
positive bacteria, Bacillus subtilis and Staphylococcus aureus (Ishikawa et
al. 2001).

Proliferation of human leukemic U937 cells has been shown to be

significantly inhibited by conditioned medium of human peripheral blood
mononuclear cells stimulated with cold-water extracts (10-800mg/mL of
medium) of dietary mushrooms, Hypsizigus mamoreus, Agrocybe
aegerite and Flammulina velutipes (Ou et al. 2005).

Ganoderma lucidum (Reishi,

Lingzhi)
A series of trials evaluating
Ganoderma lucidum in several disease
states have been carried out. The
trials evaluated effects on cancer,
Type II diabetes, coronary heart
disease, chronic hepatitis B, and
neurasthenia. Treatment with
Ganopoly for 12 weeks showed
hypoglycemic activity and produced
some anti-viral and liver protective effects in patients with chronic
hepatitis B infection. However, the same treatment regimen did not result
in any objective response in late-stage cancer patients (Zhou et al.
2005). Overall, the findings suggest that Ganopoly may have some
pharmacological activities, although clinical proof is lacking.

A double-blind, placebo-controlled, randomized, and dose-ranging study

has been carried out in men with lower urinary tract symptoms (LUTS) to

20
evaluate the safety and efficacy of an extract of Ganoderma lucidum that
shows the strongest 5-alpha-reductase inhibitory activity among the
extracts of 19 edible and medicinal mushrooms. In this trial, 88 men over
the age of 49 years who had slight-to-moderate LUTS were randomly
assigned to 12 weeks of treatment with G. lucidum extract (6mg once per
day) or placebo. The primary outcome measures were changes in the
International Prostate Symptom Score (IPSS) and variables of
uroflowmetry. Secondary outcome measures included changes in prostate
size, residual urinary volume after voiding, laboratory values, and the
reported adverse effects. G. lucidum was effective and significantly
superior to placebo for improving total IPSS with 2.1 points decreasing at
the end of treatment. No changes were observed with respect to quality
of life scores, peak urinary flow, mean urinary flow, residual urine,
prostate volume, serum prostate-specific antigen, or testosterone levels.
Overall treatment was well tolerated with no severe adverse effects
(Noguchi et al. 2008)

Ganoderma lucidum (Reishi, Lingzhi) has been reported to suppress the

invasive behaviour of breast cancer cells by inhibiting the transcription
factor NF-kappaB and to inhibit the growth of MDA-MB-231 breast cancer
cells by modulating Akt/NF-kappaB signaling (Jiang et al. 2004).

A subsequent study by the same group on the proliferation of human

estrogen-dependent (MCF-7) and estrogen-independent (MDA-MB-231)
breast cancer cells has reported that G. lucidum inhibits proliferation of
human breast cancer cells and contains biologically active compounds
with specificity against the estrogen receptor and NF-kappaB
(transcription factor) signalling (Jiang et al. 2006).

Aqueous extracts of fruiting bodies of Ganoderma lucidum, G. sinense,

and G. tsugae have been reported to have anti-tumour activities in
human breast cancer cells and immunomodulatory activities in murine
lymphocytes. In addition, it has also been suggested that the stipes of
fruiting bodies of Ganoderma species should be included in the
preparation of extracts of these fungi in order to obtain the most
comprehensive active ingredients (Yue et al. 2006).

The effect of G. lucidum on oxidative stress-induced metastatic behaviour

of poorly-invasive MCF-7 breast cancer cells has also been studied and it
has been shown that G. lucidum inhibited oxidative stress-induced
migration of MCF-7 cells by the down-regulation of mitogen activated
protein kinase (MAPK) signalling, which is involved in hormonal signalling
cascades. G. lucidum suppressed oxidative stress stimulated
phosphorylation of extracellular signal-regulated protein kinases
(Erk1/2), which resulted in the down-regulation of expression of c-fos,
21
and in the inhibition of transcription factors AP-1 and NF-kappaB. The
biological effect of G. lucidum on cell migration was mediated by the
suppression of secretion of interleukin-8 from MCF-7 cells exposed to
oxidative stress. These results suggest that G. lucidum inhibited the
oxidative stress-induced invasive behaviour of breast cancer cells by
modulating Erk1/2 signaling and could possibly be considered as an
antioxidant in adjuvant cancer therapy (Thyagarajan et al. 2006).

A further study by the same group has also shown that an extract from
green tea (GTE) increased the anti-cancer effect of G. lucidum extract
(GLE) on cell proliferation (anchorage-dependent growth) as well as
colony formation (anchorage-independent growth) of breast cancer cells.
The effect was mediated by the down-regulation of expression of the
oncogene c-myc in MDA-MB-231 cells. Although individual GTE and GLE
independently inhibited adhesion, migration and invasion of MDA-MB-231
cells, their combination demonstrated a synergistic effect, which was
mediated by the suppression of secretion of urokinase plasminogen
activator (uPA) from breast cancer cells suggesting a potential use of
combined green tea and G. lucidum extracts for the suppression of
growth and invasiveness of metastatic breast cancers (Thyagarajan et al.
2007).

The effects of Ganoderma lucidum (Basidiomycetes) polysaccharide (GL-

PS) extract on tumour volume and T(CD4+/CD8+) ratio of tumour
infiltrating lymphocytes (TILs) in breast cancer bearing mice have been
studied. The results indicated that GL-PS (100mg/kg/day) could
effectively increase the delayed type hypersensitivity response against
sRBC in BALB/c mice. Furthermore, intraperitoneal injection of this
extract in breast cancer bearing mice could increase T-cell infiltration into
the tumour. The authors concluded that GL-PS can exhibit a potent
immunomodulatory effect and may be used for potentiation of the
immune system against diseases such as cancer and other conditions in
which the immune response has been compromised (Mojadadi et al.
2006).

An alcohol extract from the spore of Ganoderma lucidum has also been
shown to inhibit the in vitro proliferation of human umbilical vein
endothelial cells and MDA-MB-231 human breast cancer cells. Further
fractionation of the alcohol extract revealed that the ethyl acetate
fraction inhibited both cell lines in a dose-dependent manner from 2 to
40mg/ml (Lu et al. 2004).

Ganoderma lucidum has also been shown to inhibit proliferation in a

dose- and time-dependent manner and induce apoptosis in human
prostate cancer cells PC-3 (Jiang et al. 2004).
22
The effects of Ganoderma lucidum on SW 480 human colorectal cancer
cells have been evaluated. A fraction containing mainly polysaccharides
(GLE-1), and a triterpenoid fraction without polysaccharides (GLE-2) were
analyzed. The data showed that both GLE-1 and GLE-2 significantly
inhibited the proliferation of SW 480 cells. The inhibitory effect of GLE-2
was much stronger than that of GLE-1. GLE-1 inhibited DNA synthesis in
the cells and reduced the formation of DPPH radicals indicating that G.
lucidum extracts inhibit proliferation of human colorectal cancer cells and
possesses antioxidant activity (Xie et al. 2006).

Aqueous extracts of the sporophores of eight mushroom species have

been assessed for their ability to prevent H2O2-induced oxidative damage
to cellular DNA using the single-cell gel electrophoresis ("Comet") assay.
The highest genoprotective effects were obtained with cold (20ºC) and
hot (100ºC) water extracts of Agaricus bisporus and Ganoderma lucidum
fruit bodies, respectively. These edible mushrooms therefore represent a
valuable source of biologically active compounds with potential for
protecting cellular DNA from oxidative damage (Rocha et al. 2002).

Protein extracts from selenium-enriched Ganoderma lucidum (Se-GLPr)

have been reported to possess strong DNA protective effects from
oxidative damage, which increased with the increase of Se content as
suggested by chemiluminescence analysis, indicating indirectly that Se
plays an important role in increasing the antioxidant activities of protein
extracts. This was confirmed by spin-trapping experiments showing that
Se-GLPr exhibited higher activities of scavenging superoxide and
hydroxyl radicals than its analog, common Ganoderma lucidum extract.
All Se-GLPr samples showed stronger activities of attenuating the
production of superoxide radical than that of hydroxyl radical (Zhao et al.
2004). Polysaccharide extracts from Se-enriched G. lucidum have also
been shown to protect DNA from hydroxyl radical oxidative damage in a
dose dependent manner (Zhao et al. 2008).

A hot water extract from Ganoderma lucidum has been shown to have an
antioxidative effect against heart toxicity in mice. Ganoderma lucidum
exhibited a dose-dependent antioxidative effect on lipid peroxidation and
superoxide scavenging activity in mouse heart homogenate. Furthermore,
this result indicated that heart damage induced by ethanol showed a
higher malonic dialdehyde level compared with heart homogenate treated
with Ganoderma lucidum. The authors concluded that this effect of
Ganoderma lucidum may protect the heart from superoxide induced
damage (Wong et al. 2004).

A human toxicological study has evaluated the consumption of Lingzhi

(Ganoderma lucidum) in a double-blinded, placebo-controlled, cross-over
23
intervention study on a range of biomarkers for human health. The study
investigated the effects of 4 weeks Lingzhi supplementation (1.44g
Lingzhi/d; equivalent to 13.2g fresh mushroom/d) on a range of
biomarkers for antioxidant status, cardiovascular disease (CHD) risk, DNA
damage, immune status, and inflammation, as well as markers of liver
and renal toxicity. No significant change in any of the biomarkers was
found. The results showed no evidence of liver, renal or DNA toxicity with
Lingzhi intake (Wachtel-Galor et al. 2004).

A recent study has investigated the anti-invasive effect of lucidenic acids

isolated from a Ganoderma lucidum strain (YK-02) against human
hepatoma carcinoma cells. The results indicated that the lucidenic acids
isolated from G. lucidum (YK-02) were anti-invasive bioactive
components on human hepatoma carcinoma cells (Weng et al. 2007).

Triterpene-enriched extracts from Ganoderma lucidum have been shown

to inhibit growth of human hepatoma Huh-7 cells via suppression of
protein kinase C, activating mitogen-activated protein kinases
(intermediates in hormonal signalling pathways) and G2-phase cell cycle
arrest. In contrast, the extracts did not inhibit growth of Chang liver cells,
a normal human liver cell line (Lin et al. 2003).

Three triterpene aldehydes, lucialdehydes A - C, from the fruiting bodies

of Ganoderma lucidum, have been shown to have cytotoxicity against
murine and human tumour cells (Lewis lung carcinoma (LLC), T-47D,
Sarcoma 180, and Meth-A tumour cell lines) (Gao et al. 2002).

Polysaccharide fractions of Ganoderma lucidum have been shown to have

potent immunomodulating effects in pre-clinical trials. A clinical study of
healthy volunteers demonstrated that G. lucidum did not affect their
immune functions. Subsequently, an open-labeled study (i.e. not double
blind or placebo controlled) aimed to evaluate the effects of water-soluble
G. lucidum polysaccharides (Ganopoly) in patients with advanced
colorectal cancer. Forty-seven patients were enrolled and treated with
Ganopoly at 5.4 g/day for 12 weeks. In 41 assessable cancer patients,
treatment with Ganopoly tended to increase mitogenic reactivity to
phytohemagglutinin (Gao et al. 2005). Larger double blind trials are
required to show if this is a real effect.

High immunomodulatory and protective effects against sarcoma 180 in

mice fed with Ling Zhi or Reishi mushroom Ganoderma lucidum (W.
Curt.: Fr.) P. Karst. (Aphyllophoromycetideae) mycelium has also been
recently reported (Rubel et al. 2008). Phenolic compounds present in
mushroom extracts from G. lucidum have also been shown to strongly

24
generate reactive oxygen species (suggesting immunomodulatory effects)
in human PBMCs and K 562 cells in vitro (Wei et al. 2008).

The polysaccharide (PS) fractions from several medicinal herbs have been
reported to have anti-ulcer effects against experimental ulcers in the rat.
The water-soluble PS fractions from Ganoderma lucidum (Reishi
mushroom) have been shown to inhibit indomethacin-induced gastric
mucosal lesions in rats. The effect of the PS fraction from G. lucidum on
the healing of gastric ulcers induced by acetic acid in the rat has
subsequently been studied. The results indicated that oral administration
of G. lucidum PS at 0.5 and 1.0g/kg for 2 weeks caused a significant
acceleration of ulcer healing by 40.1% and 55.9%, respectively. In
mechanistic studies, additional rats were treated with 10M acetic acid to
induce acute ulcers, and then treated with G. lucidum PS (1.0g/kg) for 3,
7, 10, or 14 days. Treatment with G. lucidum PS at 1.0 g/kg significantly
suppressed or restored the decreased gastric mucus levels and increased
gastric prostaglandin concentrations compared with the control group.
The results indicated that G. lucidum PS is an active component with
healing efficacy on acetic acid-induced ulcers in the rat, which may
represent a useful preparation for the prevention and treatment of peptic
ulcers (Gao et al. 2004).

Ganoderma lucidum, as well as Phellinus rimosus, Pleurotus florida and

Pleurotus pulmonaris, have been reported to have significant antioxidant
activities (Ajith and Janardhanan, 2007).

Cholesterol-lowering properties of Ganoderma lucidum have been

demonstrated in vitro, ex vivo, and in hamsters and mini-pigs. Organic
fractions containing oxygenated lanosterol derivatives inhibited
cholesterol synthesis in T9A4 hepatocytes. In hamsters, 5% Ganoderma
lucidum did not affect low density lipoprotein (LDL) but decreased total
cholesterol (TC) by 9.8%, and high density lipoprotein (HDL) by11.2%.
Ganoderma lucidum (2.5 and 5%) had effects on several faecal neutral
sterols and bile acids. In mini-pigs, 2.5% Ganoderma lucidum decreased
TC, LDL- and HDL cholesterol 20, 27, and 18%, respectively, increased
faecal cholestanol and coprostanol; and decreased cholate (Berger et al.
2004).

The hypolipidemic effect of the exo-biopolymer (EXBP) and endo-

biopolymer (ENBP) produced from a submerged mycelial culture of
Ganoderma lucidum has been investigated in dietary-induced
hyperlipidemic rats. Hypolipidemic effects were achieved in both the
EXBP- and ENBP-treated groups, however, the former proved to be more
potent than the latter. The administration of the EXBP (100mg/kg body
weight) substantially reduced the plasma total cholesterol, low-density
25
lipoprotein (LDL) cholesterol, triglyceride, phospholipid levels, and
atherogenic index by 31.0%, 39.0%, 35.4%, 28.1%, and 53.5%,
respectively, when compared to the control group. The EXBP also lowered
the liver total cholesterol, triglyceride, and phospholipid levels by 22.4%,
23.1%, and 12.9%, respectively. Furthermore, the high-density
lipoprotein (HDL) cholesterol and ratio of HDL cholesterol to total
cholesterol were significantly increased (Yang et al. 2002a).

Possible immuno-modulating effects of Ganoderma lucidum mycelium

extract (GL-M) and spore extracts on human immune cells have been
studied. Dendritic cells (DCs) are antigen-presenting cells and their role
in DC-based tumour vaccines has been well defined. The differential
effect of GL-M and GL spore extract (GL-S) on proliferation and Th1/Th2
cytokine mRNA expression of human peripheral blood mononuclear cells
(PBMCs) and monocytes has been evaluated. The effects on the
phenotypic and functional maturation of human monocyte-derived DCs
were also investigated. GL-M induced the proliferation of PBMCs and
monocytes, whereas GL-S showed a mild suppressive effect. Both
extracts stimulated Th1 and Th2 cytokine mRNA expression, but GL-M
was a relatively stronger Th1 stimulator. In contrast to GL-S, GL-M
enhanced maturation of DCs in terms of up-regulation of CD40, CD80,
and CD86, and also reduced fluorescein isothiocyanate-dextran
endocytosis. Interestingly, GL-M-treated DCs only modestly enhanced
lymphocyte proliferation in allogenic mixed lymphocyte culture with mild
enhancement in Th development. The data provide evidence that GL-M
has immuno-modulating effects on human immune cells and may be of
use as a natural adjuvant for cancer immunotherapy with dendritic cells
(Chan et al. 2005).

A polysaccharide purified from Ganoderma lucidum has also been shown

to induce gene expression changes in human dendritic cells and promotes
T helper 1 immune response in BALB/c mice (Lin et al. 2006).

Ganoderma lucidum mycelia (0.2-1.6mg/ml) have also been reported to

stimulate tumour necrosis factor-alpha (TNF-alpha) and IL (interleukin)-6
production after 8h of treatment in human whole blood. IFN (interferon)-
gamma release from human whole blood was also enhanced after 3 days
of culture with Ganoderma lucidum mycelia (0.2-1.0mg/ml). However,
Ganoderma lucidum mycelia did not potentiate nitric oxide production in
RAW264.7 cells. An electrophoretic mobility shift assay revealed that the
Ganoderma lucidum mycelia (1.6mg/ml) activated kappaB DNA binding
activity in RAW264.7 cells. These results provide supporting evidence for
the immunomodulatory effect of Ganoderma lucidum mycelia (Kuo et al.
2006).

26
An extract from Ganoderma lucidum has been reported to have apoptotic
and anti-inflammatory functions in HT-29 human colonic carcinoma cells.
Ling Zhi extract (LZE) is a herbal mushroom preparation that has been
shown to induce apoptosis anti-inflammatory action and differential
cytokine expression during induced inflammation in the human colonic
carcinoma cell line, HT-29. The extract caused no cytotoxicity in HT-29
cells at doses less than 10,000 mg/ml. Increasing concentrations reduced
prostaglandin E2 production, but increased nitric oxide production. LZE
treatment induced apoptosis by increasing the activity of caspase-3. RT-
PCR showed that LZE at a concentration of 5,000mg/ml decreased the
expression of cyclooxygenase-2 mRNA. Among 42 cytokines tested by
protein array in this study, supplementation of LZE at doses of 500 and
5,000mg/ml to HT-29 cells reduced the expression of interleukin-8,
macrophage inflammatory protein 1-delta, vascular epithelial growth
factor, and platelet-derived growth factor. These results suggest that LZE
has pro-apoptotic and anti-inflammatory functions, as well as inhibitory
effects on cytokine expression during early inflammation in colonic
carcinoma cells (Hong et al. 2004).

The potential of an Ganoderma lucidum extract as a radioprotector and

antioxidant defense against oxygen radical-mediated damage has been
studied and it was demonstrated that a hot-water extract of Ganoderma
lucidum had good radioprotective ability, as well as protection against
DNA damage induced by metal-catalyzed Fenton reactions and UV
irradiation, although the evidence was based on in vitro tests using
isolated DNA. It was also found that the water-soluble polysaccharide
isolated from the fruit body of Ganoderma lucidum was as effective as a
hot-water extract in protecting against hydroxyl radical-induced DNA
strand breaks, indicating that the polysaccharide compound is associated
with the protective properties (Kim and Kim, 1999).

The evidence for the anti-cancer effects of Ganoderma lucidum has been
reviewed (Yuen and Gohel, 2005), while the active compounds in G.
lucidum and their effects have also recently been reviewed (Boh et al.
2007).

27
Grifola frondosa (Maitake)
A fraction extracted from Grifola frondosa
(Maitake, GF-D) and its combination with
human interferon alpha-2b (IFN) has been
investigated for an inhibitory effect on
hepatitis B virus (HBV) in HepG2 2.2.15
cells (2.2.15 cells). HBV DNA and viral
antigens were analyzed by a quantitative
real-time polymerase chain reaction and
end-point titration in radioimmunoassays,
respectively. The results showed that GF-
D or IFN alone could inhibit HBV DNA in the cells with the 50% inhibitory
concentration (IC50) of 0.59mg/ml and 1399 IU/ml, respectively. The
combination of GF-D and IFN for anti-HBV activity was also evaluated and
it was found that they synergistically inhibited HBV replication in 2.2.15
cells. In combination with 0.45mg/ml GF-D, the apparent IC50 value for
IFN was 154 IU/ml. This 9-fold increase in anti-viral activity of IFN
suggested that GF-D could synergize with IFN. The results indicate that
the Grifola frondosa extract, in combination with human interferon alpha-
2b, might provide a potentially effective therapy against chronic hepatitis
B virus infections (Gu et al. 2006).

A further study by the same group has recently reported the purification
of an anti-viral protein from an extract of Grifola frondosa (Maitake)
fruiting bodies. The protein inhibited herpes simplex virus type 1 (HSV-1)
replication in vitro with an IC50 value of 4.1mg/ml and a therapeutic
index >29.3. Higher concentrations (125 and 500 mg/ml) also
significantly reduced the severity of HSV-1 induced blepharitis,
neovascularization, and stromal keratitis in a murine model. Topical
administration of the protein to the mouse cornea resulted in a significant
decrease in virus production. It was reported that the protein directly
inactivated HSV-1 while simultaneously inhibiting HSV-1 penetration into
Vero cells. The N (amino)-terminal sequence of the protein consisted of
an 11 amino acid peptide, NH2-REQDNAPCGLN-COOH that did not match
any known amino acid sequences, indicating that the protein is likely to
be a novel anti-viral protein (Gu et al. 2007).

Maitake D-fraction is a polysaccharide extracted from the Maitake

mushroom (Grifola frondosa S.F. Gray). Using normal C3H/Hej mice, its
effects on the natural immune system, including macrophages, dendritic
cells, and natural killer (NK) cells, have been investigated. NK cells attack
cells infected with pathogens such as bacteria and viruses and produce
cytokines, such as interferon-gamma (IFN-g), that can modulate natural
28
and specific immune responses. D-Fraction was administered to the mice
intraperitoneally for 3 consecutive days. Spleen cells containing
macrophages and dendritic cells were then cultured and the culture
supernatants were analyzed for IL-12. The results indicated that the D-
fraction stimulated the natural immunity related to the activation of NK
cells indirectly through IL-12 produced by macrophages and dendritic
cells, and hence administration of D-fraction to healthy individuals may
serve to prevent infection (Kodama et al. 2003).

A study has also suggested that oral administration of a submerged

cultivated G. frondosa mixture, by normal mice, may enhance host innate
immunity against foreign pathogens without eliciting an adverse
inflammatory response (Wang et al. 2008).

Anti-tumour activity induced by an extract from Grifola frondosa in a

macrophage cell line, RAW264.7 has been reported to be mediated via a
nitric oxide-mediated pathway (Sanzen et al. 2001). Similarly, an
aqueous extract from Grifola frondosa has been reported to have
immuno-modulating properties via a mechanism involving the regulation
of nitric oxide (NO) production both in vivo and in vitro (Cao et al.
2006).

The changes in the content of anti-tumour polysaccharides from Grifola

frondosa during storage have been investigated. When the mushrooms
were stored at low temperature, the content of the anti-tumour
polysaccharides showed hardly any changes, but the content decreased
markedly at higher temperature (20ºC) (Mizuno, 2000).

The photo-protective potential of exopolysaccharides (EPS) from Grifola

frondosa HB0071 has been tested in human dermal fibroblasts (HDF)
exposed to ultraviolet-A (UVA) light. It was reported that EPS had an
inhibitory effect on human interstitial collagenase (matrix
metalloproteinase, MMP-1) expression in UVA-irradiated HDF without any
significant cytotoxicity. The treatment of UVA-irradiated HDF with EPS
resulted in a dose-dependent decrease in the expression level of MMP-1
mRNA. The data suggested that EPS obtained from a mycelial culture of
Grifola frondosa HB0071 may contribute to an inhibitory action in photo-
ageing skin by reducing the MMP 1-related matrix degradation system
(Bae et al. 2005).

The biological function of GFPPS1b, a polysaccharide-peptide isolated

from cultured mycelia of Grifola frondosa GF9801, has been shown to
suppress SGC-7901 cell growth and reduce cell survival via arresting the
cell cycle in the G(2)/M phase and inducing apoptosis of tumour cells (Cui
et al. 2007).
29
The potential anti-tumour effect of beta-glucan, a polysaccharide of the
Maitake mushroom, on human prostatic cancer PC-3 cells in vitro has
been evaluated. The data showed that a bioactive beta-glucan from the
Maitake mushroom has a cytotoxic effect, presumably through oxidative
stress, on prostatic cancer cells in vitro, leading to apoptosis. This
mushroom polysaccharide may therefore have potential as a therapeutic
modality for prostate cancer (Fullerton et al. 2000). Maitake beta-
glucan has also been shown to induce hematopoietic stem cell
proliferation (Lin et al. 2007).

A beta-glucan extracted from the fruiting body of Grifola frondosa has

been reported to activate cellular immunity and expresses anti-tumour
effects, with the anti-tumour effects relating to its control of the balance
between T lymphocyte subsets Th-1 and Th-2. The fraction decreased the
activation of B cells and potentiated the activation of helper T cells,
resulting in enhanced cellular immunity. It also induced the production of
interferon (IFN)-gamma, interleukin (IL)-12 p70, and IL-18 by whole
spleen cells and lymph node cells, but suppressed that of IL-4. These
results suggest that this fraction establishes Th-1 dominance which
induces cellular immunity in the population that was Th-2 dominant due
to carcinoma (Inoue et al. 2002).

The anti-diabetic effect of an alpha-glucan (MT-alpha-glucan) from the

fruit body of Maitake mushrooms (Grifola frondosa) on KK-Ay mice (a
type 2 diabetes animal model) has been evaluated. Treatment with MT-
alpha-glucan significantly decreased body weight, level of fasting plasma
glucose, glycosylated serum protein, serum insulin, triglycerides,
cholesterol, free fatty acid and malondialdehyde content in liver.
Treatment with MT-alpha-glucan significantly increased the content of
hepatic glycogen, reduced glutathione and the activity of liver superoxide
dismutase and glutathione peroxidase. Furthermore, the insulin binding
capacity to liver crude plasma membranes increased and
histopathological changes in the pancreas were ameliorated in the
treatment group. The data suggested that MT-alpha-glucan has an anti-
diabetic effect on KK-Ay mice, which may be related to its effect on
insulin receptors by increasing insulin sensitivity and ameliorating insulin
resistance of peripheral target tissues (Lei et al. 2007).

Enhanced insulin-hypoglycaemic activity (improvement in insulin

sensitivity) has also been reported in spontaneously hypertensive rats
consuming a glycoprotein extracted from Maitake mushrooms (Preuss et
al. 2007).

The effect of Grifola frondosa total water extract on two osteoblastic cell
cultures (HOS58 and SaOS-2) has been investigated to determine if this
30
mushroom has osteo-inductive properties. The activity of alkaline
phosphate and mineralization were used as indicators for the vitality and
maturation of bone cells. The cultivation of human osteosarcoma cells
HOS58 for 5 days in the presence of an aqueous extract of G. frondosa
resulted in a significant elevation of alkaline phosphatase activity of the
cells in comparison to untreated cells. SaOS-2 cells, incubated with GFfor
21 days, showed a nearly two-fold higher mineralization than cells
cultured with a positive control, demonstrating the activity of Grifola
frondosa extract as a bone-inducing agent (Saif et al. 2007).

Plasma cholesterol concentration in rats has been shown to be reduced

by feeding of mushroom Maitake (Grifola frondosa) fiber. The results
demonstrated that mushroom fiber lowered the serum total cholesterol
level by enhancement of the hepatic low density lipoprotein (LDL)
receptor mRNA (Fukushima et al. 2001).

Maitake mushroom consumption has also been shown, in Sprague-

Dawley rats, to have the ability to alter lipid metabolism by inhibiting
both the accumulation of liver lipids and the elevation of serum lipids.
Further studies are needed to determine the mechanism of activity of
Maitake mushrooms and to establish whether their action in humans is
similar to that observed in the rat model (Kubo and Nanba, 1996). A
further study by the same group, using the same rat model system, has
also shown that consumption of dried Maitake powder (mixed with a
basic high-cholesterol rat chow) cholesterol, triglyceride and
phospholipids in the serum of rats in the Maitake-feed group were
suppressed by 0.3-0.8 times those in animals fed the basic feed.

Weights of liver and epididymal fat-pads were significantly lower (0.6-0.7

times) than those in the basic feed group, indicating that Maitake
inhibited lipid accumulation in the body. Liver lipids were also measured
and the values were found to be decreased by Maitake administration.
Measurement of the amount of total cholesterol and bile acid in faeces
showed the ratio of cholesterol-excretion had increased 1.8 fold and bile
acid-excretion 3 fold by Maitake treatment suggesting that Maitake
consumption may help to improve the lipid metabolism as it inhibits both
liver lipid and serum lipids which were increased by the ingestion of high-
fat feed (Kubo and Nanba, 1997).

Blood pressure of spontaneously hypertensive rats (SHR) has been shown

to be significantly reduced by Maitake feeding for 8 weeks, beginning at a
time when the animals were 10 weeks of age with well-established high
blood pressure. There was no difference in the plasma total and free
cholesterol, triglyceride and phospholipid levels between the Maitake fed
animals and the control (Kabir and Kimura, 1989).
31
A single oral dose of an extract of powdered Grifola frondosa, at dosage
levels of 500 and 2,000mg/kg has been given to 5 Crj:CD(SD) IGS strain
of rats of each sex for 1 day, and its toxicity was examined. The control
group was treated with water by injection. No abnormal signs were noted
in either sex of any group. No effects of powdered Grifola Frondosa were
reported in either sex by body weight measurement or necropsy finding
(Koike et al. 2003).

Hexane extracts of the cultured mycelia of Grifola frondosa have been

shown to contain ergosterol (1), ergostra-4,6,8(14),22-tetraen-3-one
(2), and 1-oleoyl-2-linoleoyl-3-palmitoylglycerol (3) and a fatty acid
fraction containing palmitic, oleic, and linoleic acids. The fatty acid
fraction and compounds 1-3 showed cyclooxygenase (COX) enzyme
inhibitory and antioxidant activities. The inhibition of COX-1 enzyme by
the fatty acid fraction and compounds 1-3 at 250mg/mL were 98, 37, 55,
and 67%, respectively. Similarly, COX-2 enzyme activity was reduced by
the fatty acid fraction and compounds 1-3 at 250mg/mL by 99, 37, 70,
and 4%, respectively. The inhibition of liposome peroxidation by the fatty
acid fraction and compounds 1 and 2 at 100 mg/mL was 79, 48, and
42%, respectively (Zhang et al. 2002).

The active compounds in Grifola frondosa (Maitake) and their effects

have recently been reviewed (Boh and Berovic, 2007).

Hericium erinaceum (Pom Pom, Lion’s Mane)

Anti-dementia compounds have recently
been reported from the mushroom
Hericium erinaceum (Kawagishi, 2007;
Kawagishi and Zhuang, 2008). Hericium
erinaceum extracts have also been
reported to induce neurite outgrowth from
neural (NG108-15) cells. Maximum
stimulation of neurite outgrowth was
recorded with mycelial extracts, and the
least stimulation was observed with an
oven-dried fruit body extract. Aqueous
extracts of H. erinaceus therefore contain neuroactive compounds that
stimulate neurite outgrowth in vitro suggesting some value in the
treatment of neurodegenerative disease (Wong et al. 2007)

Immuno-regulatory functions of Hericium erinaceum have been

demonstrated in an aqueous extract by a stimulation of inducible nitric
oxide gene expression followed by nitric oxide production in macrophages

32
via enhancement of activation of the transcription factor, NF-kappaB (Son
et al. 2006).

Polysaccharides from Hericium erinaceum and Hericium laciniatum have

been extracted from culture broth and the polysaccharide components
were mainly glucose in H. erinaceus and galactose in H. laciniatum. Both
polysaccharides had significant anti-artificial pulmonary metastatic
tumour effects in mice with the polysaccharide from H. erinaceus being
more effective than that from H. laciniatum. However, both of the
polysaccharides enhanced the increase of T cells and macrophages
(immuno-enhancing activity) (Wang et al. 2001).

The hypolipidemic effect of exo-polymers produced in submerged

mycelial cultures of Hericium erinaceus (HE), Auricularia auricula judae
(AA), Flammulina velutipes (FV), Phellinus pini (PP), and Grifola frondosa
(GF) has been investigated in dietary-induced hyperlipidemic rats. The
animals were administered with exo-polymers at the level of 100mg/kg
body weight daily for four weeks. A hypolipidemic effect was achieved in
all the experimental groups, however, HE exo-polymer proved to be the
most potent, significantly reducing plasma triglyceride (28.9%), total
cholesterol (29.7%), low-density lipoprotein (LDL) cholesterol (39.6%),
phospholipid (16.0%), and liver total cholesterol (28.9%) levels,
compared to the saline administered (control) group. The results
demonstrated the potential of Hericium erinaceus exo-polymer in treating
hyperlipidemia in dietary-induced hyperlipidemic rat (Yang et al.
2002c; Yang et al. 2003).

A methanol extract of the fruiting bodies of Hericium erinaceus has been

fed to rats and shown to result in a significantly lower elevation rate of
blood glucose level than control rats. The effects on blood glucose, serum
triglyceride and total cholesterol levels were more significant in the rats
fed daily with the Hericium erinaceus extract at doses of 100mg/kg body
weight rather than 20mg/kg body weight (Wang et al. 2005).

Hypsizigus marmoreus (Buna-shimeji)

Anti-tubercular activity and an inhibitory
effect on Epstein-Barr virus activation of
sterols and polyisoprenepolyols from an
edible mushroom, Hypsizigus
marmoreus (Buna-shimeji) have been
reported. Seven sterols and eight
polyisoprenepolyols, isolated from the
non-saponifiable lipid fraction of the

33
dichloromethane extract of Hypsizigus marmoreus, have been tested for
their anti-tubercular activity against Mycobacterium tuberculosis strain
H37Rv using the Microplate Alamar Blue Assay (MABA). Six of the sterols
and two of the polyisoprenepolyols showed a minimum inhibitory
concentration (MIC) in the range of 1-51 mg/ml, while the others were
inactive. The seven sterols and three polyisoprenepolyols were further
evaluated for their inhibitory effects on Epstein-Barr virus early antigen
(EBV-EA) activation induced by the tumour promoter 12-O-
tetradecanoylphorbol-13-acetate (TPA) in Raji cells. Two of the sterols
showed a potent inhibitory effect while preserving the high viability of the
Raji cells (Akihisa et al. 2005).

Anti-tumour-promoting activity has been found in methanol and ethyl

acetate extracts of the mushroom 'Buna-shimeji', Hypsizigus marmoreus
(Tricholomataceae). From the active fractions of the extracts, two sterols,
ergosterol and ergosterol peroxide, were isolated. The isolates showed
inhibitory activity against 12-O-tetradecanoylphorbol-13-acetate induced
ear inflammation in mice, and ergosterol markedly inhibited tumour
promotion in a two-stage carcinogenesis experiment (Yasukawa et al.
1994).

Aqueous and methanol extracts of Hypsizigus marmoreus have been

tested against allogeneic tumour, solid sarcoma 180 and syngeneic
tumour and Meth A fibrosarcoma. The aqueous extract was highly active
in inhibiting growth of solid sarcoma 180, but not as effective for Meth A
fibrosarcoma. Fractionation of anti-tumour substances of the aqueous
extract isolated four polysaccharides. Chemical analysis revealed one of
them to be beta-(1-3)-glucan with a significant inhibitory effect against
tumour-growth of sarcoma 180 (Ikekawa et al. 1992).

Chloroform extracts of the fruit bodies of Hypsizigus marmoreus have

been shown to exhibit moderate cytotoxicity towards cultured human
colon carcinoma (HT-29), human breast carcinoma (MCF-7) and human
hepatoblastoma (HepG-2) cell lines (Xu et al. 2007).

Anti-proliferative activities of fractions of Hypsizigus marmoreus have

been examined using HepG2 cells in vitro. The methanol extract of
H.marmoreus markedly induced anti-proliferative activity and an active
compound from this mushroom has been identified as hypsiziprenol A9.
Hypsiziprenol A9 inhibited cell proliferation in a time- and concentration-
dependent manner by up to 80% on HepG2 cells by inducing arrest of the
G1 phase. Hypsiziprenol A9 also decreased expression of phosphorylated
retinoblastoma protein (ppRb), cyclin D1, and cyclin E in a dose-
dependent manner. The results suggested that hypsiziprenol A9 can

34
inhibit the growth of HepG2 cells through inducing G1 phase cell cycle
arrest due to the inhibition of pRb phosphorylation (Chang et al. 2004).

A novel ribonuclease, from fresh fruiting bodies of Hypsizigus

marmoreus,with anti-proliferative activity against the L1210 leukemia cell
line has also been purified (Guan et al. 2007). A thermostable
ribosome-inactivating protein with a molecular weight of 20kDa, isolated
from fruiting bodies of Hypsizigus marmoreus has also been shown to
have anti-proliferative activity against mouse leukemia cells and human
leukemia and hepatoma cells (Lam and Ng, 2001).

The antioxidant effects of Hypsizygus marmoreus have been studied for

peroxyl and alkoxyl radicals by ordinary, non-tumour-bearing and
tumour-bearing mice. Oral administration of the fruit body of H.
marmoreus exhibited potent anti-tumour or cancer-preventive effects and
caused a significant decrease in lipid peroxide levels, which were
determined as thiobarbituric acid reactive substances. These results
showed that the intake of H. marmoreus fruit body could induce an
antioxidant effect, and the increase of antioxidant activity in the plasma
of tumour-bearing mice was an important mechanism in cancer
prevention. It was also suggested that the mushroom might play a role in
the decrease of lipid peroxides through antioxidant activity induction
(Matsuzawa, 2006).

Proliferation of human leukemic U937 cells has been shown to be

significantly inhibited by conditioned medium of human peripheral blood
mononuclear cells stimulated with cold-water extracts (10-800 mg/mL
of medium) of Hypsizigus mamoreus, Agrocybe aegerite and Flammulina
velutipes (Ou et al. 2005).

The isolation of a collagen-binding protein from Hypsizigus marmoreus,

which inhibits the Lewis lung carcinoma cell adhesion to type IV collagen
has been reported. A type IV collagen-binding protein of 23kDa was
isolated from the mushroom, Hypsizigus marmoreus. This protein, HM
23, bound to type IV and type I collagens and gelatin, and to much lesser
extent to fibronectin, but not to laminin or bovine serum albumin. The
adhesion of Lewis lung carcinoma cells was inhibited when the type IV
collagen substratum was pretreated with HM 23 (Tsuchida et al. 1995).

Bunashimeji-related hypersensitivity pneumonitis has been reported in

workers who cultivate this mushroom in indoor facilities in Japan. An
evaluation of protective measures concluded that complete cessation was
the best treatment for hypersensitivity pneumonitis. The use of a mask
was ineffective for patients with a high serum Krebs von der Lungen-6
(KL-6), surfactant protein-D (SP-D) concentration and severe ground-
35
glass opacity on chest high-resolution computed tomography. Initial
treatment with oral prednisolone was recommended for patients with
high levels of total cell counts in bronchoalveolar lavage fluid (Tsushima
et al. 2006).

Lentinula edodes (Shiitake)

Extracts from Lentinula edodes (Shiitake) have
been widely reported to have anti-tumour
activity. However, this activity has been shown to
be host-mediated and not by direct cytotoxic
activity to cancer cells. A very recent study
(Israilides et al. 2008) has now demonstrated
cytotoxic and cell growth inhibitory (cytostatic)
effects of aqueous extracts of the mushroom on
the MCF-7 human breast adenocarcinoma cell
line. The effect was demonstrated with fruit body
and mycelial extracts, the difference being that
there was no significant suppression on normal
cells with the latter. Furthermore, mycelial extracts did not induce any
cytostatic effect in both cancer and normal cell lines based on a DNA
synthesis assay. The significant suppression of the proliferation of cancer
cells was reflected by the comparatively low IC50 values and the
simultaneous higher respective values on normal fibroblast cells. In
addition to the direct inhibition of the proliferation of human breast
cancer cells in vitro, the Lentinula edodes extract had immunostimulatory
properties in terms of mitogenic and co-mitogenic activity in vitro.

The effects of diets containing dry powdered Shiitake mushroom on

frequency of azoxymethane-induced colon aberrant crypt foci (ACF) and
intestinal tumours in male Sprague-Dawley rats have been studied.
Pregnant rat dams and their progeny were fed AIN-93G diets containing
casein (20%; control diet) or casein supplemented with Shiitake (1% or
4% wt/wt). Casein- and 1% Shiitake-fed rats exhibited identical growth
curves, whereas those fed the 4% Shiitake diet were of slightly reduced
body weight. The 4% Shiitake diet elicited increased active energy
expenditure and reduced adiposity of rats. Small bowel and colon
tumours and colon ACF were evaluated in the male progeny at 18 weeks
after azoxymethane treatment. Aberrant crypt foci and tumours were
most prevalent in the mid and distal regions of the colon. Shiitake intake
had no effect on the relative incidence of tumours in the colon or small
intestine (duodenum). Consumption of 1% Shiitake stimulated growth of
invasive adenocarcinomas in the mid colon and favoured a non-significant
36
increase in median frequency of ACF in this same region. In contrast,
Shiitake at 4% intake elicited a reduction in colon tumour multiplicity.
The authors suggested a stimulatory action of 1% Shiitake on rat colon
tumourigenesis which is puzzling as the data were not statistically
significant. However, the inhibitory actions of 4% Shiitake mushroom on
the indices of rat colon tumourigenesis where statistically validated
(Frank et al. 2006).

Shiitake extracts have been dispersed with lecithin micelles to prepare

superfine particles (0.05 to 0.2 microns in diameter) of beta-1,3-glucan
(micellary mushroom extracts). When mice were fed with these micelles
of beta-glucan (0.75mg/day/mouse, smaller amounts of beta-glucan),
the number of lymphocytes yielded by the small intestine increased by up
to 40% and tumour cytotoxicity against P815 cells and cytokine
production was also augmented, suggesting that smaller amounts of
micellary beta-glucan might be useful for the potentiation of intestinal
immunity (Shen et al. 2007).

The effects of protein-bound polysaccharides (A-PBP and L-PBP),

extracted from the mycelia of Agaricus blazei and Lentinus edodes, on
serum cholesterol and body weight have been investigated in 90 female
volunteers. The data demonstrated a weight-controlling and
hypolipidemic effect of both A-PBP and L-PBP via a mechanism involving
absorption of cholesterol (Kweon et al. 2002).

The effect of Shiitake (Lentinus edodes, LE) and autolyzed- (fermented-)

Shiitake (autolyzed-LE) on blood pressure and serum fat levels of
spontaneously hypertensive rats (SHR) have been studied. The animals
of the autolyzed-LE group showed significantly lower blood pressure
compared to the control or LE group. The serum levels of total cholesterol
(TC), triglyceride and phospholipid of the groups fed with LE and
autolyzed-LE were lower than those of the control group, and atherogenic
index [(TC-HDL-C)/HDL-C] improved significantly in 21 days. It was
suggested that the serum TC decline is the action of eritadenine that is
contained in the Shiitake mushroom. An inhibitory activity of the
angiotensin I-converting enzyme (ACE) was compared between of LE and
autolyzed-LE. Autolyzed-LE showed higher inhibitory activity than LE
against the ACE. The results suggested that the hypotensive action of
autolyzed-LE was due to concomitant ACE inhibitory activities of peptides
and gamma-aminobutyric acid contained in higher amounts during the
autolysis of LE (Watanabe et al. 2002).

The changes in the content of an anti-tumour polysaccharide from

Lentinus edodes (lentinan) during storage have been investigated. When
the mushrooms were stored at low temperature, the content of their anti-
37
tumour polysaccharides showed hardly any changes, but the content
decreased markedly at higher temperature (20ºC) (Mizuno, 2000).

The Shiitake mushroom (Lentinus edodes) contains the

hypocholesterolemic agent eritadenine, 2(R),3(R)-dihydroxy-4-(9-
adenyl)-butyric acid. A study has recently been conducted to quantify the
amount of the cholesterol reducing agent eritadenine in Shiitake
mushrooms. The amounts of eritadenine in the fruit bodies of four
different shiitake mushrooms, Le-1, Le-2, Le-A, and Le-B, were
investigated. Methanol extraction was used to recover as much
eritadenine as possible from the fungal cells, and enzymes that degrade
the fungal cell walls were also used to elucidate if the extraction could be
further enhanced. The Shiitake strains under investigation exhibited up to
10-fold higher levels of eritadenine than previously reported for other
Shiitake strains. Pre-treatment of mushrooms with hydrolytic enzymes
before methanol extraction resulted in an insignificant increase in the
amount of eritadenine released. The results suggested the potential for
delivery of therapeutic amounts of eritadenine from the ingestion of
extracts or dried concentrates of Shiitake mushroom strains (Enman et
al. 2007).

Plasma cholesterol concentration in rats has been shown to be reduced

by feeding of mushroom Shiitake (Lentinus edodes) fiber. The results
demonstrated that mushroom fiber lowered the serum total cholesterol
level by enhancement of the hepatic low density lipoprotein (LDL)
receptor mRNA (Fukushima et al. 2001).

Methanol and water extracts from Lentinus edodes have been shown to
have antioxidant activity against lipid peroxidation of rat brain
homogenate. The antioxidant activity against lipid peroxidation was found
to correlate with the phenolic content in different sub-fractions of the
mushroom extracts (Cheung and Cheung, 2005).

The effect of heat treatment on the changes in the overall antioxidant

activity and polyphenolic compounds of Shiitake extract has been
investigated. Raw Shiitake was heated at 100 and 121ºC for 15 or 30 min
using an autoclave. After heat treatment, the free and bound
polyphenolics and flavonoids in the mushroom extracts were analyzed.
2,2-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical and
1,1-diphenyl-2-picrythydrazyl (DPPH) radical scavenging activities were
measured to evaluate antioxidant activity of the extracts. The
polyphenolic contents and antioxidant activities in the extracts increased
as heating temperature and time increased. The free polyphenolic
content in the extract heated at 121ºC for 30 min was increased by 1.9-
fold compared to that in the extract from the raw sample. The ABTS and
38
DPPH radical scavenging activities were increased by 2.0-fold and 2.2-
fold compared to the raw sample, respectively. The data showed that
heat treatment significantly enhanced the overall antioxidant activities of
Shiitake mushrooms (Choi et al. 2006a).

High-molecular-weight polysaccharides (HMWP), including lentinan, in

Shiitake mushrooms may promote human health. A study has been
conducted to determine if management protocols influence the HMWP of
Shiitake (Lentinula edodes (Berk.) Pegler) mushrooms. The results
indicated that measuring the total carbohydrate content of water-
extractable, ethanol-insoluble polysaccharides was a simple way to
estimate HMWP. The results also indicated that log-grown Shiitake
contained more HMWP than did substrate-grown Shiitake. Among log-
grown Shiitake, both mushroom strain and tree species influenced HMWP
content. The results suggested that there is considerable variation among
Shiitake mushrooms in HMWP content and that production protocols
influenced the HMWP content of mushrooms (Brauer et al. 2002).

Supplemental amounts of a polysaccharide/oligosaccharide complex

obtained from a Shiitake mushroom extract have been evaluated for the
ability to lower the prostate-specific antigen level in patients (n=62) with
prostate cancer. The data showed that the Shiitake mushroom extract
alone was ineffective in the treatment of clinical prostate cancer (White et
al. 2002). A high genistein, Shiitake mushroom extract has also been
reported to have anti-tumour effects on prostate cancer (Hackman et al.
2001).

An ethyl acetate fraction from Shiitake (Lentinus edodes) mushrooms has

been investigated using two human breast carcinoma cell lines (MDA-MB-
453 and MCF-7), one human non-malignant breast epithelial cell line
(MCF-10F), and two myeloma cell lines (RPMI-8226 and IM-9).
Concentration-dependent anti-proliferative effects of the fraction were
observed in all cell lines. Approximately 50mg/L of the fraction induced
apoptosis in 50% of the population of four human tumour cell lines and
the fraction-induced apoptosis may have been mediated through the pro-
apoptotic bax protein which was up-regulated. Cell cycle analysis
revealed that the fraction induced cell cycle arrest by a significant
decrease of the S phase, which was associated with the induction of cdk
inhibitors (p21) and the suppression of cdk4 and cyclin D1 activity.
Compared to malignant tumour cells, non-malignant cells were less
sensitive to the fraction for the suppression of cell growth and regulation
of bax, p21, cyclin D1, and cdk4 expression. A 51% anti-proliferative
effect occurred at the highest concentration of the fraction (800mg/L).
The data suggest that inhibition of growth in tumour cells by the Shiitake

39
mushroom extract may result from an induction of apoptosis (Fang et al.
2006).

The isolation and characterization of an anti-tumour polysaccharide, KS-

2, extracted from culture mycelia of Lentinus edodes has also been
reported. KS-2 suppressed the growth of EHRLICH as well as Sarcoma-
180 tumours in mice when given either orally or intraperitoneally (Fujii et
al. 1978).

Extracts from fermentation broth and mycelium of 15 strains of Lentinus

edodes have been shown to be active against gram positive and gram
negative bacteria, yeasts and mycelial fungi, including dermatophytes
and phytopathogens. The strains differed by the set of the organisms
susceptible to the action of the extracts. Strains of L. edodes combining
marked anti-bacterial properties and high yields of water soluble
polysaccharides were screened. The active compounds were detected by
preparative thin layer chromatography. Two were identified with UV- and
mass spectrometry as lentinamycin B and erytadenine (lentinacin).
Lentinamycin B was found to be the main component responsible for the
anti-bacterial activity of the L. edodes strains (Soboleva et al. 2006).

The anti-microbial activity of the culture fluid of Lentinus edodes

mycelium grown in submerged liquid culture has been demonstrated
against Streptococcus pyogenes, Staphylococcus aureus and Bacillus
megaterium. The substance responsible for the activity was heat-stable
and was suggested to be lenthionine, an anti-bacterial and anti-fungal
sulphur-containing compound (Hatvani, 2001).

A study on the action of lentinan (extracted from Shiitake mushrooms

(Lentinus edodes) has been conducted using murine lymphoma (K36)
cells in a AKR mouse model. Further investigation on the effectiveness of
the extracted lentinan was then performed using human colon-carcinoma
cell lines in mice. Six established human colon-carcinoma cell lines
segregated into three groups of different degrees of differentiation were
used in this study. One group was not fed (control) and the second group
was prefed with lentinan for 7 days prior to inoculations with the cancer
cells. The size of the tumours that developed was rated after 1 month.
Significant regression in tumour formation was observed in prefed mice
compared to control (unfed) mice when K36 or human colon-carcinoma
cells were used. Significant reductions in the size of the tumours were
observed in mice prefed with lentinan. Follow-up investigation proceeded
with the use of nude mice (athymic). Lymphocytes extracted from AKR
mice prefed with lentinan for 7 days were inoculated into the nude mice.
This was then followed by inoculation of the human colon-carcinoma cell
lines into these mice. Much smaller tumours were formed in nude mice
40
inoculated with lymphocytes, in contrast to the larger tumour formed in
nude mice without lymphocyte inoculation. The study concluded that the
anti-tumour property of lentinan was maintained with oral administration.
In addition, "primed" lymphocytes, when given passively to immuno-
deficient mice, were able to retard the development of tumours in these
mice (Ng and Yap, 2002).

The hypoglycemic effect of exo-polymers (EPs) produced from

submerged mycelial cultures of five varieties of mushrooms on
streptozotocin (STZ)-induced diabetic rats have been investigated. The
five experimental groups were fed with EPs (50mg/kg body weight) for 7
days. Significant reductions in plasma glucose, total cholesterol (TC), and
triglyceride (TG) levels were observed in rats fed with Lentinus edodes
and Cordyceps militaris EPs. Plasma glucose and TC were also reduced by
administration of Phellinus linteus EPs, but the TG level was not changed
significantly. The EPs of the three mushroom species also demonstrated a
marked reduction in the level of plasma glutamate-pyruvate
transaminase (GPT). The result demonstrated the hypoglycemic activity
of EPs of three mushroom varieties in STZ-induced diabetic rats and
suggests some potential in the control of diabetes mellitus (Kim et al.
2001).

A subsequent study by the same group, using higher concentrations

(200mg/kg body weight in streptozotocin-induced diabetic rats) of exo-
polymers from a submerged mycelial culture of Lentinus edodes has
shown that the administration of the exo-polymer reduced the plasma
glucose level by as much as 21.5%, and increased plasma insulin by
22.1% compared to the control group. It was also shown to lower the
plasma total cholesterol and triglyceride levels by 25.1 and 44.5%,
respectively (Yang et al. 2002b).

Three anti-bacterial compounds extracted by chloroform, ethylacetate or

water from dried Shiitake mushrooms (Lentinus edodes) have been
reported which possessed efficient anti-bacterial activities against
Streptococcus spp., Actinomyces spp., Lactobacillus spp., Prevotella spp.,
and Porphyromonas spp. of oral origin. In contrast, other general
bacteria, such as Enterococcus spp., Staphylococcus spp., Escherichia
spp., Bacillus spp., and Candida spp. were relatively resistant to these
compounds. The anti-bacterial activity of chloroform extracts and
ethylacetate extracts were relatively heat-stable, while the water extract
was heat-labile (Hirasawa et al. 1999).

The action of the juice of Shiitake mushrooms (L. edodes) at a

concentration of 5% from the volume of the nutrient medium was found
to produce a pronounced anti-microbial effect with respect to Escherichia
41
coli O-114, Staphylococcus aureus, Enterococcus faecalis, Candida
albicans and to stimulate the growth of E. coli M-17. Bifidobacteria and
Lactobacteria exhibited resistance to the action of L. edodes juice
(Kuznetsov et al. 2005).

Shiitake dermatitis after the ingestion of raw Shiitake mushrooms has

been reported, primarily in Japan, and it has been suggested that this
dermatitis may be photosensitive as nearly half of the patients studied
developed the dermatitis on skin exposed to sunlight (Hanada and
Hashimoto, 1998). A study in Korea has also reported dermatitis effects,
but in contrast to the previous reports in Japan, cases with Shiitake
dermatitis occurred after eating boiled or cooked Shiitake mushrooms
suggesting that a non-thermolabile factor/component may be involved
(Ha et al. 2003).

A study has been conducted with 10 people where each participant

ingested 4g of Shiitake powder daily for 10 weeks (trial 1), and the
protocol was repeated in the same subjects after 3 to 6 months (trial 2).
Gastrointestinal symptoms coincided with eosinophilia in two subjects.
Symptoms and eosinophilia resolved after discontinuing Shiitake
ingestion. The authors reported that daily ingestion of Shiitake mushroom
powder in five of 10 healthy persons provoked blood eosinophilia,
increased eosinophil granule proteins in serum and stool, and increased
gastrointestinal symptoms (Levy et al. 1998).

A single oral dose of an extract of cultured Lentinus edodes mycelia, at

dosage levels of 500 and 2,000mg/kg has been given to 5 Crj:CD(SD)
IGS strain of rats of each sex for 1 day, and its toxicity was examined.
The control group was treated with water by injection. No abnormal signs
were noted in either sex of any group. No effects of Lentinus edodes
mycelia were reported in either sex by body weight measurement or
necropsy finding (Koike et al. 2002a). A follow on study by the same
authors that extended the treatments for 28 days reported no effects in
either sex by body weight measurement, food consumption
measurement, urinalysis, ophthalmological examination, hematological
examination, blood chemical analysis, necropsy finding, organ weight
measurement, or histopathological examination (Koike et al. 2002b).

42
Lyophyllum connatum
A new ergothioneine derivative, beta-
hydroxyergothioneine has been isolated from the
mushroom Lyophyllum connatum. Ergothioneine,
N-hydroxy-N',N'-dimethylurea, and connatin (N-
hydroxy-N',N'-dimethylcitrulline) were also
isolated. All the compounds displayed the ability
to scavenge free radicals, based on a 1,1-
diphenyl-2-picrylhydrazyl (DPPH) radical
scavenging assay. Structural determination,
including the absolute stereochemistry of beta-
hydroxyergothioneine, was achieved by
spectroscopic analysis and X-ray crystallography.
The radical scavenging activity of beta-
hydroxyergothioneine was almost the same as that of ergothioneine.
Beta-hydroxyergothioneine showed the greatest protective activity
against carbon tetrachloride-induced injury in primary culture
hepatocytes (Kimura et al. 2005).

Phellinus linteus
Ethanol extracts of Phellinus linteus have
been shown to have antioxidant activities
comparable to Vitamin C in scavenging the
stable free radical 1,1-diphenyl-2-
picrylhyrazyl (DPPH). The extracts also
inhibited lipid peroxidation (LPO) in a
concentration-dependent manner. The
study also reported anti-angiogenic
activities of Phellinus linteus (Song et al.
2003).

Phellinus linteus has been reported to sensitise apoptosis induced by

doxorubicin (an anti-cancer drug) in prostate cancer LNCaP cells
suggesting that Phellinus linteus may have therapeutic potential to
augment the magnitude of apoptosis induced by anti-prostate cancer
drugs (Collins et al. 2006). Phellinus linteus has also been shown to
mediate cell-cycle arrest at a low concentration and apoptosis in response
to a high dose in mouse and human lung cancer cells (Guo et al. 2007).

Phellinus linteus has been reported to contain constituents that exhibit

potent anti-tumour effects through activation of immune cells. A recent
study in mice has reported that boiling water soluble fractions from
43
mycelium of P.linteus contain anti-allergic and immuno-potentiating
properties (Inagaki et al. 2005).

An acidic polysaccharide from Phellinus linteus has been shown to

markedly inhibit melanoma cell metastasis in mice, and directly inhibit
cancer cell adhesion to, and invasion through, the extracellular matrix,
but that it had no direct effect on cancer cell growth. In addition, the
authors reported that PL increased macrophage NO production. These
results suggest that Phellinus linteus has two anti-metastatic functions -
it acts as an immuno-potentiator and as a direct inhibitor of cancer cell
adhesion (Han et al. 2006).

An extract from Phellinus linteus has been shown to have anti-

inflammatory activity (Kim et al. 2004) via mediation of heme
oxygenase-1 in an in vitro inflammation (macrophage) model (Kim et al.
2006).

The effect of a mushroom extract of Phellinus linteus on non-cancerous

prostate cells using an experimentally developed rat benign prostatic
hyperplasia model has been studied. The results showed that prostate
weight increased significantly by 37% owing to treatment with the
mushroom extract, and in particular, the stromal component of the
prostate increased significantly by 80%. A suppression of transforming
growth factor-beta1 expression by 56% was observed with the
mushroom extract treatment. It was found that the mushroom extract
enlarged the prostate and therefore administration of Phellinus linteus
extract should be considered carefully by those with an enlarged prostate
(Shibata et al. 2005).

Phellinus linteus has also been shown to suppress growth, angiogenesis

and invasive behaviour of breast cancer cells (Sliva et al. 2008).

Phellinus robustus
It has been reported that melanins from the
medicinal mushroom Phellinus robustus have
high antioxidant and geno-protective
properties (Babitskaya et al. 2007).

44
Pholiota nameko (Nameko)
Hypersensitivity pneumonitis to spores
of Pholiota nameko has been reported
in a mushroom farmer, although
separation from the antigen along with
corticosteroid therapy, resulted in the
symptoms and inflammatory effects
quickly subsiding (Inage et al.
1996).

Pleurotus eryngii
The effects of Pleurotus eryngii extracts (PEX) on
bone metabolism have been studied. PEX
treatment showed an increase in the alkaline
phosphatase activity of osteoblasts and in the
osteocalcin mRNA expression from primary
osteoblasts. PEX also increased the expression of
the Runx2 gene, and the secretion of
osteoprotegerin from the osteoblasts showed
marked increases after treatment with PEX. In
vivo studies, using rats with ovariectomy-induced
osteoporosis revealed that PEX alleviated the decrease in the trabecular
bone mineral density (Kim et al. 2006).

The ergothioneine content of mushrooms has been reported to be in the

range of 0.4-2.0mg/g (dry weight). The white Agaricus bisporus
contained the least ergothioneine and portabellas (brown) contained the
highest within the varieties of A. bisporus studied. The specialty
mushrooms tested (Lentinus edodes, Pleurotus ostreatus, P. eryngii,
Grifola frondosa) all contained a statistically significant greater amount of
ergothioneine compared to A. bisporus, however, no significant difference
was found between the specialty mushrooms (Dubost et al. 2006).

The antioxidative potency of commercially available mushrooms in

Taiwan has been studied. The order of inhibitory activity of mushroom
extracts on oxidation in emulsion system was Agaricus bisporus >
Hypsizigus marmoreus > Volvariella volvacea > Flammulina velutipes >
Pleurotus eryngii > Pleurotus ostreatus > Hericium erinaceus > Lentinula

45
edodes. In a thermal oxidative stability test, using lard, the order of
antioxidative activity of the mushroom extracts showed similar
tendencies, except for the extract of Lentinula edodes (Fui et al. 2002).

Mycelial extracts (ethyl acetate and 70% ethanol) of 20 higher

Basidiomycetes edible or medicinal mushrooms and culture media
extracts (ethyl acetate and Butan-1-ol) have been evaluated for in vitro
anti-inflammatory activity using the cyclooxygenase-1 and -2 enzymes
(COX-1 and -2). In general, 70% ethanolic extracts showed lower
inhibitory activity against both enzymes compared to ethyl acetate
extracts. Of the mushrooms tested, the ethyl acetate extracts of
Ganoderma applanata, Naematoloma sublateritium, Pleurotus eryngii,
and P. salmoneostramineus showed the highest COX-2 inhibitory effects
compared to COX-1 inhibition. Of the culture media tested in this study,
only the ethyl acetate extracts of the culture medium of Agrocybe
cylindracea exhibited high inhibition of the COX-2 enzyme (Elgorashi et
al. 2008). Ceramide from Agrocybe aegerita has also been reported to
inhibit the cyclooxygenase enzymes, COX-1 and -2 (Diyabalanage et al.
2008).

Hypersensitivity pneumonitis induced by Pleurotus eryngii spores has

been reported in a worker in an Eringi (Pleurotus eryngii) mushroom
factory who had worked there for 6 years. Chest radiography showed
diffuse fine nodular shadows. Chest computed tomography demonstrated
centrilobular nodules and increased attenuation in both lungs. The patient
suffered from hypoxemia while breathing room air. The lymphocyte count
in the bronchoalveolar lavage fluid was increased, and transbronchial
lung biopsy specimens showed lymphocyte alveolitis with epithelioid cell
granulomas in the alveolar spaces. After admission, the patient's
symptoms improved rapidly without medication. However, on his return
to work, fever and hypoxemia appeared again. The lymphocyte
stimulating test was positive against extracts of Eringi spores. Precipitins
against the extracts of Eringi spores were detected by the double
immunodiffusion test. The diagnosis was hypersensitivity pneumonitis
(HP) caused by Eringi spores. In Japan, more than 30 cases of HP
induced by mushroom spores have been reported and therefore this is an
occupational health and safety issue, related to air quality in mushroom
factories that needs to be addressed. The symptoms appear to improve
without medication (Miyazaki et al. 2003).

46
Pleurotus ferulae
Ethanol and hot water extracts of
Pleurotus ferulae have been shown to
have anti-tumourigenic properties in
human cervical cancer and human lung
cancer cell lines. When A549, SiHa and
HeLa cells were incubated with different
concentrations of ethanol and hot water
extracts, the ethanol extracts showed
strong cytotoxicity against A549 cells at
concentrations over 10 mg/mL and against SiHa and HeLa cells at
concentrations over 40 mg/mL. The ethanol extracts were the most
prominent anti-tumour agents (of those studied) toward A549 human
lung cancer cells (Choi et al. 2004a).

Pleurotus Ostreatus (Oyster mushrooms)

The effect of Oyster mushrooms on reduction of
blood glucose, cholesterol and triglycerides in
diabetic patients has been evaluated in a clinical
investigation of 89 subjects. Mushroom
consumption significantly reduced systolic and
diastolic blood pressure, lowered plasma
glucose, total cholesterol and triglycerides
significantly, whereas there was no significant
change in body weight. There were no
deleterious effects on liver or kidney function
(Khatun et al. 2007).

A hypo-cholesterolemic effect has been shown

with Oyster mushrooms (Pleurotus Ostreatus) in rats with Streptozotocin-
induced diabetes. Oyster mushroom (4% dry oyster mushroom fruit
body) lowered cholesterol content by more than 60% in the liver
although it did not significantly affect either the serum triacylglycerol
level or the content in liver (Bobek et al. 1991). Similar results have
been observed in rats with a hereditary hypersensitivity to dietary
cholesterol (Bobek et al. 1990). The hypo-cholesterolemic effects of
Oyster mushrooms has been demonstrated to be dose-dependent (Bobek
et al. 1997). A similar hypocholesterolemic effect of the oyster
mushroom (Pleurotus ostreatus) was also observed in hamsters (Bobek
et al. 1993) and in rabbits (Bobek and Galbavy, 1999).

47
The addition of 4% dried whole oyster mushroom (Pleurotus Ostreatus)
to the diet of Wistar rats has been reported to have led to a reduced level
of serum and liver cholesterol at the end of the 10th week of the
experiment. The level of serum triacylglycerols was not influenced by the
mushroom, but was significantly reduced by 13% in liver. The decrease
in serum cholesterol level was a consequence of the decreased
cholesterol concentration in very-low-density lipoproteins (VLDL) and in
low-density lipoproteins (LDL). The content of cholesterol in high-density
lipoproteins (HDL) was not influenced by the mushroom. Dietary
Pleurotus ostreatus increased the fractional turnover rate of LDL (by
28%) and HDL (by 31%) as determined by the analysis of decay curves
of 125I-labelled lipoproteins. The increase in the rate of LDL and HDL
catabolism is one of the mechanisms which mediates the
hypocholesterolemic effect of mushrooms in the rat model (Bobek et al.
1993).

Feeding of 5% powder of the fruiting bodies of the Oyster mushroom

(Pleurotus ostreatus) to hyper-cholesterolaemic rats reduced their plasma
total cholesterol by ~28%, low-density lipoprotein-cholesterol by ~55%,
triglyceride by ~34%, non-esterified fatty acid by ~30% and total liver
cholesterol levels by > 34%, with a concurrent increase in plasma high-
density lipoprotein-cholesterol concentration of > 21%. However, these
effects were not observed in mushroom-fed normocholesterolaemic rats.
Mushroom feeding significantly increased plasma fatty acid unsaturation
in both normo- and hypercholesterolaemic rats, while plasma total
antioxidant status was significantly decreased in mushroom-fed
hypercholesterolaemic rats, concomitant with a decrease in plasma total
cholesterol. The study concluded that 5% P. ostreatus supplementation
provides health benefits, at least partially, by acting on the atherogenic
lipid profile in the hyper-cholesterolaemic condition (Hossain et al.
2003).

The effects of pleuran, a beta-glucan isolated from Pleurotus ostreatus,

have been studied in a model of acute colitis in rats. Pleuran was given
either as a 2% food component or as a 0.44% pleuran hydrogel drink
over 4 weeks. Colitis was induced by intraluminal instillation of 4% acetic
acid and after 48h the extent of colonic damage and several biochemical
parameters were examined. Pleuran supplementation both in food and in
drinking fluid significantly decreased the disposition to colitis. The
enhanced activity of myeloperoxidase in the inflamed colonic segment
was reduced by pleuran diets, reflecting decreased neutrophil infiltration.
The mechanism of the described protective effect of pleuran is not yet
clear, but the authors suggest that the pleuran-enhanced antioxidant

48
defence of the colonic wall against the inflammatory attack maybe a
factor (Bobek et al. 2001).

In vivo injection of three water-soluble proteoglycan fractions from

Pleurotus ostreatus mycelia, which had polysaccharide to protein ratios
14.2, 26.4 and 18.3 respectively, into Sarcoma-180-bearing mice
decreased the number of tumour cells and cell cycle analysis showed that
most of the cells were found to be arrested in pre-G(0)/G(1) phase of the
cell cycle. All of the three proteoglycans elevated mouse natural killer
(NK) cell cytotoxicity and stimulated macrophages to produce nitric
oxide. Fourier transform infra red (FTIR) spectra suggested the presence
of a beta-glycosidic bond in all the fractions (Sarangi et al. 2006).

Anti-proliferative and pro-apoptotic activities of fractions of Pleurotus

ostreatus have been evaluated in HT-29 colon cancer cells in vitro. A hot-
water-soluble fraction of the mycelium of the liquid cultured mushroom
was partially isolated and chemically characterized as a low-molecular-
weight alpha-glucan. This low-molecular-weight alpha-glucan possessed
anti-tumourigenic properties, and demonstrated its direct effect on colon
cancer cell proliferation via induction of apoptosis - programmed cell
death (Lavi et al. 2006).

A dimeric lectin isolated from fresh fruiting bodies of Pleurotus ostreatus

has been shown to possess potent anti-tumour activity in mice bearing
sarcoma S-180 and hepatoma H-22. Survival in these mice was
prolonged and body weight increase reduced after lectin treatment (Wang
et al. 2000).

Pleurotus pulmonarius
Hypoglycaemic activity of an aqueous
extract of Pleurotus pulmonarius in
alloxan-induced diabetic mice has been
reported. Pleurotus pulmonarius
extract was administrated orally at
doses of 250, 500, and 1,000mg/kg to
separate groups of mice (normal and
alloxan-treated mice), and serum
glucose and body weight were
measured. In the separate group of
mice, an oral glucose tolerance test
was carried out. Acute oral toxicity data showed no mortality in the
normal mice up to 5,000mg/kg, while oral administration of extracts
reduced the serum glucose level in alloxan-treated diabetic mice at all the

49
doses tested after acute and chronic (28 days) administration. The
extract also showed increased glucose tolerance in both normal and
diabetic mice. The data suggest that the extract possesses
hypoglycaemic activity (Badole et al. 2006).

In a subsequent study by the same group, the interaction of an aqueous

extract of Pleurotus pulmonarius with acarbose on serum glucose levels,
and on an oral glucose-tolerance test in alloxan induced diabetic mice
was studied. The anti-hyperglycaemic effects of aqueous extract and
acarbose alone were similar but combination treatment of the Pleurotus
pulmonarius extract with acarbose produced a more synergistic anti-
hyperglycaemic effect than either agent alone (Badole and Bodhankar,
2007).

Podaxis pistillaris
Anti-bacterial components of the mushroom Podaxis
pistillaris have recently been reported. Podaxis pistillaris
(Podaxales, Podaxaceae, Basidiomycetes) was found to
exhibit anti-bacterial activity against Staphylococcus
aureus, Micrococcus flavus, Bacillus subtilis, Proteus
mirabilis, Serratia marcescens and Escherichia coli. In a
culture medium of P. pistillaris, three
epidithiodiketopiperazines were identified by activity-
guided isolation. Based on spectral data their identity was
established as epicorazine A(1), epicorazine B(2) and
epicorazine C (3, antibiotic F 3822), which have not
previously been reported as constituents of P. pistillaris (Al-Fatimi et al.
2006).

Schizophyllum commune (Split Gills Mushroom)

A case of mucoid impaction of the bronchi due
to a hypersensitivity reaction to the
monokaryotic mycelium of Schizophyllum
commune has been reported. The patient was
hospitalized because of mild asthma attacks,
persistent cough, peripheral eosinophilia, and
"gloved finger" shadows on a chest
roentgenogram. Cultures of the mucous plugs
and sputum samples yielded white, felt-like
mycelial colonies that were later identified as
the monokaryotic mycelium of S. commune by

50
use of mating tests with established monokaryotic and dikaryotic strains
of S. commune. The results of tests for serum antibody to S. commune
cytosol antigen were positive. Bronchoscopies were effective in removing
the mucous plugs and relieving the patient's symptoms. The authors
suggested that the monokaryotic mycelium of S. commune should be
considered as one of the fungi that can cause hypersensitivity-related
lung diseases (Amitani et al. 1996). The incidence of Schizophyllum
commune related effects in patients suffering from diseases of the nasal
sinuses also appears to be on the increase (Buzina et al. 2003).

A lectin from the split gill mushroom Schizophyllum commune has been
shown to exhibit potent mitogenic activity toward mouse splenocytes,
anti-proliferative activity toward tumour cell lines, and inhibitory activity
toward HIV-1 reverse transcriptase, but did not possess any anti-fungal
activity (Han et al. 2005).

Tremella fuciformis (White Wood Ear, White Jelly Leaf)

Tremella fuciformis has been shown to have

hypo-cholesterolemic properties in rats
(Cheung, 1996b).

Tremetes (Coriolus) versicolor (Turkey tail, Yunzhi)

Yunzhi (Coriolus versicolor) has been
reported to modulate various
immunological functions in vitro, in vivo,
and in human clinical trials, while
Danshen (Salvia miltiorrhiza) has been
shown to benefit the circulatory system
by its vasodilating activity. A clinical trial
has been carried out to evaluate the
immunomodulatory effects of Yunzhi-Danshen capsules in post-treatment
breast cancer patients. Eighty-two patients with breast cancer were
recruited to take Yunzhi and Danshen capsules with the data showing
that the percentage and the absolute counts of B-lymphocytes were
significantly elevated in patients with breast cancer after taking Yunzhi-
Danshen capsules, while plasma sIL-2R concentration was significantly
51
decreased (Wong et al. 2005). The significance of these findings is not
yet clear.

A polysaccharopeptide from the Turkey Tail fungus Trametes (=Coriolus)

versicolor has been reported to be capable of inhibiting human
immunodeficiency virus type 1 (HIV-1) reverse transcriptase and
protease, the two enzymes of paramount importance to the life cycle of
the HIV. The polysaccharopeptide inhibited other proteases including
trypsin, chymotrypsin, proteinase K, subtilisin, and elastase to a lesser
extent. The anti-HIV enzyme and immuno-stimulatory activities of the
mushroom polysaccharopeptide make it an interesting potential candidate
for the therapeutic treatment of AIDS (Tzi et al. 2006), although clearly
further studies are required to confirm such effects. A recent report has
also suggested that the culture (harvest) duration affects the
immunomodulatory and anti-cancer effects of polysaccharopeptides
derived from Coriolus versicolor (Lee et al. 2006)

Coriolus versicolor (CV), also known as Trametes versicolor, has been

evaluated for its cytotoxic activities on a B-cell lymphoma (Raji) and two
human promyelocytic leukemia (HL-60, NB-4) cell lines. The results
demonstrated that CV extract at 50 to 800mg/ml dose-dependently
suppressed the proliferation of Raji, NB-4, and HL-60 cells by more than
90%. The extract however did not exert any significant cytotoxic effect
on the normal liver cell line WRL when compared with a
chemotherapeutic anti-cancer drug, mitomycin C, confirming the tumour-
selective cytotoxicity. Nucleosome production in HL-60, NB-4 and Raji
cells was significantly increased by 3.6-, 3.6- and 5.6-fold respectively
upon the treatment of CV extract, while no significant nucleosome
production was detected in extract-treated WRL cells. The CV extract was
found to selectively and dose-dependently inhibit the proliferation of
lymphoma and leukemic cells possibly via an apoptosis-dependent
pathway (Lau et al. 2004).

Coriolus versicolor polysaccharide (CVP) extracted from C. versicolor dry

fruit bodies by hot-water extraction and ethanol precipitation has been
evaluated against four human liver cancer (7703, HepG2, 7721, PLC) and
four human breast cancer (Bcap37, ZR75-30, MCF-7, T-47D) cell lines
using a cytotoxicity assay. The results showed that the CVP inhibited the
proliferation of 7703, Bcap37, T-47D in low concentration and also
inhibited the proliferation of MCF-7 and ZR75-30, but at high
concentrations. The CVP did not inhibit the proliferation of HepG2, 7721,
PLC and human normal liver cell line (WRL). The CVP was found to
selectively inhibit the proliferation of human liver cancer and human
breast cancer (Zhou et al. 2007).

52
The effect of dietary supplementation with a protein-bound
polysaccharide (PSP)-containing extract derived from mycelia of Coriolus
versicolor on in vitro and in vivo indices of immune function of young and
old C57BL/6NIA mice has been studied. Young (5 month) and old (23
month) mice were fed purified diets containing 0%, 0.1%, 0.5% or 1.0%
PSP for 1 month after which time indices of immune function were
measured. PSP supplementation had no significant effect on mitogenic
response to concanavalin A (Con A), phytohemagglutinin (PHA) or
lipopolysaccharide (LPS), or on the production of interleukin (IL)-1, IL-2,
IL-4 and prostaglandin E-2 (PGE(2)). Of the in vivo indices of immune
function tested, old mice fed 1.0% PSP had significantly higher delayed-
type hypersensitivity (DTH) response than those fed 0% PSP. No
significant effect of PSP was observed on the DTH response of young
mice. The antibody response to sheep red blood cells was not significantly
influenced by PSP in young or old mice, suggesting that the PSP-
containing extract from mycelia of Coriolus versicolor might have a
modest immuno-enhancing effect in aged mice, but not in young mice
(Wu et al. 1998).

Toth et al. have recently reported the inhibition of intestinal cancer by a

hot water extract of the Coriolus versicolor (Turkey Tail) mushroom in
C57bl/6j-Apc(Min) mice (Toth et al. 2007).

A highly N-methylated cyclic heptapeptide, isolated from the mushroom

Coriolus versicolor, has been shown to have an inhibitory effect on fat
accumulation by 3T3-L1 murine adipocytes (EC50 = 0.02mg/mL)
(Shimokawa et al. 2008).

Volvariella volvacea (Straw Mushroom)

Methanol and water extracts from Volvariella
volvacea have been shown to have antioxidant
activity against lipid peroxidation of rat brain
homogenate. The ethyl acetate sub-fraction of the
methanol extract of V. volvacea was found to have
comparable antioxidant activity to caffeic acid
against the oxidation of human low-density
lipoprotein (LDL). The antioxidant activities
against lipid peroxidation were found to correlate
with the phenolic content in different sub-fractions
of the mushroom extracts (Cheung and Cheung, 2005).

A report on the fractionation of extracts of the edible mushroom,

Volvariella volvacea, has shown the isolation of two heterocyclic

53
carboxylic acids, namely pyridine-3-carboxylic acid [nicotinic acid] and
pyrazole-3(5)-carboxylic acid and four steroidal metabolites ergosterol,
5-dihydroergosterol, ergosterol peroxide, and cerevisterol. In light of the
structural similarity of pyrazole-3(5)-carboxylic acid to pyrazole-3-
carboxylic acids, which act as agonists for nicotinic acid receptors, the
levels of pyridine-3-carboxylic acid and pyrazole-3(5)-carboxylic acid
were estimated in V. volvacea and two other edible mushrooms, namely
Agaricus bisporus and Calocybe indica. Significant levels of pyridine-3-
carboxylic acid (nicotinic acid) were found in C. indica, and pyrazole-3(5)-
carboxylic acid was found in abundance in A. bisporus. Correlations have
been suggested between the occurrence of these compounds in
mushrooms and consumption as well as beneficial health effects
(Mallavadhani et al. 2006).

Volvariella volvacea has been reported to produce a hypotensive

response in animals. An aqueous extract of the mushroom (SME) was
prepared and given through intravenous injections to normotensive rats.
An i.v. injection of SME produced a hypotensive effect in rats with an
ED50 of 25mg dry weight/kg body weight. SME did not increase urinary
excretion or sodium diuresis. It produced positive chronotropic and
inotropic effects on isolated right atria and induced contraction of isolated
tail artery strips. This latter contractile response was inhibited by
antagonists of serotonin and alpha-adrenoceptor, ketanserin and
phentolamine respectively. Partial purification using dialysis and liquid
chromatography revealed that the hypotensive active substances had
molecular weights between 8,000 and 12,000 daltons. These substances
were heat stable and resistant to trypsin digestion (Chiu et al. 1995).

Volvariella volvacea has also been shown to have hypo-cholesterolemic

properties (Cheung, 1996a; Cheung, 1998).

54
Wild Edible Fungi
While fungi are a good source of digestible
proteins and fibre, are low in fat and energy and
make a useful contribution to vitamin and
mineral intake, nevertheless, there are some
safety concerns with wild fungi. Edible species
might be mistaken for poisonous ones, high
heavy-metal concentrations (in some regions
and countries) in wild edible fungi (WEF) are a
known source of chronic poisoning and the
consumption of WEF can contribute markedly to
the radiocaesium intake of human subjects
(reported for the UK). Some regions of Europe
have a strong WEF tradition, particularly eastern
Europe. Only one-third of adults consume fungi
(cultivated species and WEF) throughout the UK;
the average intake of fungi in the UK being estimated to be 0.12 kg fresh
weight per capita per year. At least eighty-two species of wild fungi are
recorded as being consumed in the UK, although certain species (e.g.
chanterelle (Cantharellus cibarius), cep (Boletus edulis), Oyster
mushroom (Pleurotus ostreatus)) are favoured over others. Although wild
edible fungi are not essential components in the daily diet, they have
been reported to be a nutritionally-valuable addition to the range of
vegetables consumed (de Roman et al. 2006).

In contrast to the above study, a recent analysis of a large variety (7

different families) of wild edible mushrooms in Greece has shown high
mineral contents and low contents of heavy metals (Pb, Cd and As)
suggesting that the mushroom species studied can be consumed without
a risk to health (Ouzouni et al. 2007)

55