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Current Research in Food Science 5 (2022) 2190–2203
Available online 5 November 2022
2665-9271/© 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Critical review on chemical compositions and health-promoting effects of
mushroom Agaricus blazei Murill
Kaiyuan Huang
a
,
b
, Hesham R. El-Seedi
c
,
d
, Baojun Xu
a
,
*
a
Food Science and Technology Program, Department of Life Sciences, BNU-HKBU United International College, Zhuhai, China
b
Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, China
c
Pharmacognosy Group, Department of Pharmaceutical Biosciences, Uppsala University, Biomedical Centre, Box 591, SE, 751 24, Uppsala, Sweden
d
International Research Center for Food Nutrition and Safety, Jiangsu University, Zhenjiang, 212013, China
ARTICLE INFO
Keywords:
Agaricus blazei Murill
Phytochemical constituents
Immunoregulation
Inammation
Pharmacological uses
ABSTRACT
Agaricus blazei Murrill (AbM) is a medical mushroom which has huge potential commercial value with various
health-promoting functions. However, the chemical composition and therapeutic mechanisms of AbM have not
been concluded systematically yet. Thus, this study aims to comprehensively summarize the phytochemical
proles and thoroughly characterize the health promotion effects such as the antitumor and antidiabetic impact
of AbM in in vivo and in vitro. The AbM consists of abundant bioactive substances; polysaccharides, lipids
including ergosterol, sterols, proteins, vitamin B, C and D, and phenolic compounds. Several studies have claimed
that Agaricus blazei Murrill polysaccharides (AbMP) had immunoregulation, anti-inammatory, hep-
atoprotective, and antitumor function both in vivo and in vitro. Meanwhile, AbM extracts were thought to cure
diabetes and bacterial infection, exhibiting anticarcinogenic and antimutagenic functions. But some principles
behind health-promoting effects have not been claried. Additionally, AbM related clinical trials are limited and
further discovery need to be conducted. Therefore, this paper has concluded the health promotion impact with
corresponding mechanisms of AbM and indicated its potential medical usage as functional food in the future.
1. Introduction
Mushrooms have been considered a functional and nutritional food
worldwide due to their low calories and high content of bioactive
components, proteins, lipids, minerals, and vitamins. In the early 21st
century, higher basidiomycetes as medical mushrooms have attracted
much attention due to their nutritional and pharmacological properties
(Delmanto et al., 2001). Agaricus blazei Murill (AbM), scientic name
Cyperus mushroom, is an edible Basidiomycetes mushroom, which was
rst discovered in Florida, U.S.A., in 1944 and is known as Hime-
matsutake in Japan and Ji-songrong in China (Wu and Wang, 2000).
AbM was rst brought to Japan in 1965; with the maturity of articial
cultivation technology, it has been widely used in tea and food, and later
gained a reputation for its excellent medical value and biochemical
properties since the 1980s. In many countries, such as in Brazil, people
use the extracts as natural therapy and tea, which is widely accepted as a
new method to prevent and treat cancer (Delmanto et al., 2001; Mizuno,
2002).
Murrill (1945) comprehensively described the morphological
characteristics of AbM; the cap of was initially hemispherical, up to
1–1.5 cm thick in the centre, 7–9 cm broad, margin glabrous, at in the
middle, and its color ranged from white to light gray or dark
reddish-brown. Then it becomes convex with a diameter of 6–11 cm, and
the surface was covered with lamentous bers which would form small
scales when matured. The spores exhibited dark brown color, smooth,
broadly oval, and spherical, about 5 ×4
μ
m, without bud holes, and the
hyphae had no lock-like association (Figure 1). Tian and Ren (2009)
further claimed that AbM cultivation would be benecial to the devel-
opment of circular agriculture and conservation agriculture. As a
saprophytic fungus, AbM can be planted entirely with crop straw, while
operations such as bagging and sterilization are not in need. On the other
hand, picking process of AbM is convenient with a high input-output
ratio since it be refrigerated for freshness or dried and dehydrated for
transportation or storage. Moreover, the optimum temperature ranged
from 22 to 26◦C, and 60–70% moisture content without light and a
6–7.5 pH value should be maintained to the culture material.
In China, the concept of homology of medicine and food occupies an
important position in traditional Chinese medicine theory (Zhou and Xu,
* Corresponding author. Jintong Road, Tangjiawan, Zhuhai, Guangdong, 519087, China.
E-mail address: baojunxu@uic.edu.cn (B. Xu).
Contents lists available at ScienceDirect
Current Research in Food Science
journal homepage: www.sciencedirect.com/journal/current-research-in-food-science
https://doi.org/10.1016/j.crfs.2022.10.029
Received 27 July 2022; Received in revised form 15 October 2022; Accepted 30 October 2022
Current Research in Food Science 5 (2022) 2190–2203
2191
2021). On the other hand, AbM had a relatively short history of con-
sumption in China, which was rst introduced to Fujian Province in
China from Japan in 1992. Since then, Chinese scientists are more
enthusiastic on the research of health promotion function of the mush-
room in the past few decades. AbM has been evidenced by several in vivo
and in vitro studies since 21st and utilized medicinally to treat various
diseases including diabetes, arteriosclerosis, chronic hepatitis, hyper-
lipidemia, and cancer (Hetland et al., 2008). Additionally, AbM was
capable of regulating cellular immunity, antioxidant, antibacterial and
anti-inammatory responses (Firenzuoli et al., 2008). Wang et al. (2014)
and Halpern (2007) proved that AbM could signicantly strengthen the
immune system by improving the activities of natural killer cells and
increasing the number of white blood cells. And it was also generally
believed that AbM extracts exhibited antioxidant, hepatoprotective and
anti-viral activities (Sorimachi et al., 2001). Thus, nutrition composi-
tions of AbM aroused heated attention among researchers.
Ohno et al. (2001) systematically investigated the constituents of
AbM and found it mainly consisted of polysaccharides,
(1–3)-β-D-β-glucans with side branches of a (1–6)-β-backbone, gener-
ating considerable immunomodulatory and antitumor effects by acti-
vation of cytotoxic macrophages, helper T cells, and NK cells, promotion
of T cell differentiation, and activation of the alternative complement
pathway. The authors suggested high molecular weight of triple helical
structure may be responsible for immunopotentiation activity, but few
data can indicate was independent of any specic ordered structure.
More recently, Menezes et al. (2022) demonstrated that AbM poly-
saccharides (AbMP) had good rheological properties, and which can be
used to develop pharmaceutical formulations with antioxidant activity
and low cytotoxicity to human neutrophils. Furthermore,
sulfated-modied AbMP were claimed to increase their anti-HIV phar-
macological activity, holding the promise for a future biomedical
treatment of HIV/AIDS (Zhao et al., 2021). Besides, ergosterol, rstly
isolated by Takaku et al. (2001) from AbM lipid was demonstrated its
signicant tumor growth inhibition ability in vivo study without side
effects. In addition, vitamin B
1
, B
2
, B
9,
B
12
, vitamin C, vitamin D and
niacin could also be found in the AbM fruit body (Mizuno, 1995; R´
ozsa
et al., 2019). The scientists suggested that these pharmacological nu-
trients can be isolated from different parts of the mushroom. In partic-
ular, fruit bodies, pure culture mycelia, and culture ltrates were
considered fundamental parts that were rich in bioactive nutrients
extracted by microwaving, organic solvents or hot water extraction and
centrifuging (Sun et al., 2011; Shen et al., 2001; Li, 2020). An increasing
number of scientists are trying to obtain active metabolites from mycelia
through deep fermentation to obtain cheaper preparations for immu-
noregulation or disease treatment (Firenzuoli et al., 2008). Some com-
positions may not isolate from the mushroom; however, they still
expressed the bioactive functions overall. Sorimachi et al. (2001) re-
ported that AbM mycelium also revealed cytopathic inhibition effects of
western equine encephalitis virus on cultured VERO cells. Fan et al.
(2006) documented the efcacy of the AbM compound combined with
chemotherapy for gastric cancer. They conrmed that white cells and
CD4
+
/CD8
+
signicantly increased after treatment, which indicated
that AbM could relieve clinical symptoms with the combination of
chemotherapy. Beyond that, some AbM-based medical products have
been developed and evidenced by clinical trials. AndoSan™ was one of
the clinical anti-inammatory medicines which can treat Crohn’s dis-
ease (CD) by reducing pro-inammatory cytokines in patients (Ther-
kelsen et al., 2016). Although existing clinical studies on the
pharmacological effects of AbM were relatively limited, it has been
widely used and considered complementary and alternative medicine in
worldwide (Wang et al., 2013).
Existing reviews have rarely concluded for their nutrition composi-
tion and potential therapeutic effects of AbM. Thus, this critical review
aims to comprehensively summarize AbM chemical proles with their
nutritional value; the existing bioactivities of AbM. On the other hand,
some mechanisms were thoroughly summarized for the further clinical
application and bioactive principal discovery. Also, the impact of food
processing of AbM were compared. The review collected the previous
studies across the past 40 years via screening of databases such as
PubMed, Google Scholar, ResearchGate, etc., using the corresponding
keywords. The tables and gures were generated by Excel (version
16.62), XMind and BioRender.
2. Nutritional values of AbM
AbM as a functional food, which enjoyed great popularity for
numerous high-quality bioactive nutrient components. In general, active
metabolites can be isolated from AbM fruiting bodies, pure cultured
mycelium, submerged fermentation culture from mycelium and culture
ltrate. There are differences in the content of biologically active sub-
stances in different parts of AbM (mycelia and fruit body) and between
treatment methods (dry powder, hydroalcoholic extracts and fresh
AbM). The detail gross compositions of mushrooms and the contents of
different parts were collected in Table 1. The AbM comprised vibrant
carbohydrates (59.42 g/100g), such as oligosaccharides, mannans,
xylose,
α
-glucans, and β-glucans and mainly contained four mono-
saccharides: glucose, galactose, mannose and fucose (Carneiro et al.,
2013; Kim et al., 2005; Liu et al. 2022; OHNO et al., 2001; Mizuno et al.,
1990). Moreover, AbMP including AbEXP1-a (Li, 2020), AbMP-II-
α
and
AbMP-II-β (Shen et al., 2001), FI
0
-a-β, FA-1-a-⍺, FA-a-β (Mizuno et al.,
1990), agaritine (Akiyama et al., 2011) could be extracted from AbM.
Uronic acid as an acid glucan accounted for about 28.19%, which may
potentially affect their bioactive activities (Wu et al., 2014). On the
other hand, ergosterol (73–90 mg/100 g) was isolated from AbM lipid
fractions by Takaku et al. (2001). Ergosterol, as a precursor of vitamin
D
2
(1.95–3.68 mg/100g), was usually present in AbM mushrooms at
higher levels than other crops (R´
ozsa et al., 2019; Vieira Junior et al.,
2021). Normally, fat accounted for 1.82 g/100 g (2–8%) of the AbM
(Carneiro et al., 2013; Firenzuoli et al., 2008). In addition, ber (3–32%)
and vitamin C (11–21 mg/100g) were rich in common edible mush-
rooms including AbM (Horm and Ohga 2008; Takaku et al., 2001).
Vitamin B
1
, B
2
, B
9,
B
12
and niacin could be found in AbM, especially in
fruit organisms (Mizuno, 1995; R´
ozsa et al., 2019). Moreover, Sun
(2007) claimed that the mycelium and fruiting body of AbM contain
various essential trace elements, especially zinc (Zn) and selenium (Se).
Hemagglutinin (ABL) as a lectin that can be extracted from the fruiting
body of fresh AbM (Kawagishi et al. 2001).
Fig. 1. Agaricus blazei Murill (AbM) fresh and dry samples were purchased in
Taobao on June 29, 2022 in Yunnan province, China.
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2192
3. Flavor and taste compositions
Chen (1986) claimed that mannitol was the taste-active compounds
in mushroom sugars and 7.94 g/100g can be detected in AbM by Car-
neiro et al. (2013). On the other hand, the soluble sugars such as glucose
(83.59 %), fructose (1.04 %) contained in mushrooms contribute to the
sweet taste (Carneiro et al., 2013; Liu et al., 2017). Umami or palatable
taste in mushroom contributed by the synergistic effect of 5′-nucleotides
(5.51 mg/g in AbM mycelia) with MSG-like components. And aspartic
(0.50 mg/g) and glutamic acids (not detected in AbM) were mono-
sodium glutamate-like (MSG-like) components, which gave the most
typical mushroom taste in AbM (Chang et al., 2001). In addition, Yang
et al. (2022) suggested umami amino acids and nucleotides potentially
enhanced the salty intensity of NaCl. With the increasing demand of
low-salt foods, the synergistic effect of umami and salty could satisfy the
need of consumers who tend to acquire a low-salt diet. Thus, umami
fungus including AbM could be potentially used as the ingredient for
future functional food development.
4. Bioactivities of AbM
4.1. Immunoregulation and Anti-fatigue effect
The spleen and thymus are essential central and peripheral immune
organs of the body. As the largest secondary lymphoid organ in the body,
the spleen could regulate T- and B-cell responses to these antigenic
targets in the blood (Lewis et al., 2019). On the other hand, the cytokines
such as tumor necrosis factor-
α
(TNF-
α
), Interleukin-6 (IL-6), and
Interleukin-1β (IL-1β) were generated by immune cells, including mac-
rophages and lymphocytes for the immunoregulation of human (Sun,
2017). Li and Wei (2017) indicated that the AbMP had signicant
bioactive effects on fatigue through above-mentioned indicators. Based
on their research in Figure 2, the mice were divided into four groups,
and the three groups received low, medium, and high gradient doses
(100, 150, and 200 mg/kg BW per day, respectively) of AbMP. The other
group was served the same amount of normal saline as the control group.
After seven days of adaptive feeding, mouse thymus and spleen indices;
serum IL-6, TNF-
α
; T lymphocytes: CD4
+
and CD8
+
; were determined
separately. Li and Wei (2017) demonstrated that AbMP signicantly (p<
0.05) promoted the growth of the spleen and thymus in adult mice, and
the thymus and spleen indexes of each dose group were increased to a
certain extent compared with the control group. Furthermore, medium
and high doses (150 and 200 mg/kg per day, respectively) of AbMP
increased signicantly (p <0.05) the secretion of TNF-
α
and IL-6 to
regulate the immune function of the body. Similarly, CD4
+
and CD8
+
as
two vital T lymphocytes in adaptive immunity, and their number of cells
signicantly increased when treated with medium and high levels of
AbMP. Thus, the authors declared that AbMP could signicantly
improve mice’s humoral, cellular, and non-specic immunity. Apart
from that, superoxide dismutase (SOD) and malondialdehyde (MDA) are
essential indexes to evaluate exercise fatigue (Li and Wei, 2017). SOD is
the most important free radical scavenger in the body, and MDA is the
nal product of lipid peroxidation when free radicals act on lipids in
living organisms, which reects the degree of cell damage and can
scavenge free radicals in the body (Peng et al., 2011). Thus, the authors
also measured the concentration of SOD and MDA in mice after con-
sumption of AbMP. The results exhibited that the concentration of SOD
and MDA in the serum of the AbM polysaccharide dose groups of 150
and 200 mg/kg BW concentration was signicantly different from that
of the control group (p <0.05). SOD is the leading free radical scavenger
in the body, and the concentration of SOD in the serum of mice was
signicantly increased from 368.25 ±24.23 to 478.62 ±43.53 U/mg,
which had signicant antifatigue activity. In the experiment, the MDA
concentrations were signicantly lower than the control group (17.37 ±
2.46 and 23.54 ±1.47
μ
mol/mg, respectively), which indicated that
AbM extracts could reduce the process of free radical oxidation and
Table 1
The relative proportion of nutritional compounds and phytochemicals in Agar-
icus blazei Murill (AbM).
Composition Content (mean ±SD,
percentage, or content
range)
Reference
Carbohydrates 59.42 ±1.86 g/100g (DP);
121.2 ±9.20
μ
g/mg (FE)*
Carneiro et al. (2013);
Carvajal et al., 2011
Fat 1.82 ±0.03 g/100 g (DP);
2–8 % (FW)
Carneiro et al. (2013);
Firenzuoli et al.
(2008)
Proteins 31.29 ±1.85 g/100g (DP);
2–40 % (FW)
Carneiro et al. (2013);
Firenzuoli et al.
(2008)
Total sugars 66.91 ±7.58 g/100 g (DP) Carneiro et al. (2013)
Total soluble sugar (g/
100g)
1.72 ±0.17 (FW) Sun et al. (2011);
Crude Ash 5–7 %; 7.47 ±0.04 (DP) Takaku et al. (2001);
Carneiro et al. (2013)
Fiber 3–32 % (FW) Takaku et al. (2001);
Firenzuoli et al.
(2008)
Moisture 85–90 % (FW) Takaku et al. (2001);
Firenzuoli et al.
(2008)
Total tocopherols 124.25 ±31.30
μ
g/100g
(DP)
Carneiro et al. (2013)
Total phenolic and related
compounds
0.77 ±0.12 mg/100g (DP) R´
ozsa et al. (2019)
Uronic acid 28.19 ±1.39% (Freeze
drying)
Wu et al. (2014)
Mannose 49.12–80.71 mg/100g
(FW); 0.59 % (DP)
Sun et al. (2011); Liu
et al. (2017)
Trehalose 23.9 ±0.24 mg/g (MY);
5.74 ±0.70 mg/100g (DP)
Chang et al. (2001);
Carneiro et al. (2013)
Arabitol 31.4 ±0.68 mg/g (MY) Chang et al. (2001)
Galactose 5.46 % (DP); 210.43 ±8.22
mg/100g (FW)
Liu et al. (2017); Sun
et al. (2011)
Glucose 403.19–852.23 mg/100 g
(FW); 83.59 % (DP)
Sun et al. (2011); Liu
et al. (2017)
Ergosterol 73.00–90.17 mg/100g (DP) R´
ozsa et al. (2019)
Mannitol 7.94 mg/100g (DP) Carneiro et al. (2013)
Flavonoids 1.8 ±0.16
μ
g/mg (FE)* Carvajal et al. (2011)
Fructose 0.27 ±0.02 g/100 g (DP);
1.04 % (DP)
Carneiro et al. (2013);
Liu et al. (2017)
L-Aspartic acid 0.50 ±0.06 mg/g (MY) Chang et al. (2001)
L-Glutamic acid nd (MY) Chang et al. (2001)
p-hydroxybenzoic acid 0.64 ±0.09 mg/100g (DP) Carneiro et al. (2013)
Vanillic acid nd (DP) Carneiro et al. (2013)
Trans-p-coumaric acid 0.08 ±0.02 mg/100g (DP) Carneiro et al. (2013)
Gallic acid 4.50 ±0.10
μ
g/mg (FE) * Carvajal et al. (2011)
Syringic acid 5.70 ±0.10
μ
g/mg (FE) * Carvajal et al. (2011)
Pyrogallo acid 3.5 ±0.10
μ
g/mg (FE) * Carvajal et al. (2011)
Cinnamic acid 0.05 ±0.01 mg/100g (DP) Carneiro et al. (2013)
Viatmin B1 381–1151
μ
g/100g (DP) R´
ozsa et al. (2019)
Viatmin B2 3183–5616
μ
g/100g (DP) R´
ozsa et al. (2019)
Viatmin B9 291–671
μ
g/100g (DP) R´
ozsa et al. (2019)
Viatmin B12 463–906
μ
g/100g (DP) R´
ozsa et al. (2019)
Viatmin C 11.00–21.67 mg/100g (DP) R´
ozsa et al. (2019)
Vitamin D2 1.95–3.68 mg/100g (DP) Vieira Junior et al.
(2021)
Vitamin E 124.25 ±31.30
μ
g/100 g
(DP)
Carneiro et al. (2013)
Flavor 5′-nucleotides 5.51 ±1.35 mg/g (MY) Chang et al. (2001)
a: MY, Content in mycelia
of AbM
b: DP, dry powder of AbM
c: FW, Fresh weight of AbM
d: FE, AbM fruit body after
hydroalcoholic extract
e: *, of glucose equivalents
f: nd, not detected.
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2193
resist the fatigue of the body.
Additionally, peripheral fatigue is caused by the depletion of blood
sugar and tissue glycogen due to the accumulation of lactic acid and
ammonia metagarite. Glycolysis is the primary energy metabolism
pathway during intense exercise, while blood lactate and hematuria
nitrogen are the primary metabolites that indicate physical fatigue (Li
et al., 2008; Li and Wei, 2017). Duan et al. (2003) found that the content
of crude polysaccharides in the fermentation mycelium of AbM was
much higher than that in the body. AbEXP1-a was a water-soluble
polysaccharide which was isolated and puried by Li (2020) from
crude extracellular AbMP, suggesting its antifatigue activity in vivo test.
Fifty 35 g mice ingested 0.2 mL/kg BW of AbM polysaccharide per day,
and their antifatigue indicators were measured after three weeks. The
hematuria nitrogen and blood lactate levels after quantitative exercises
in the test group were 7.84 ±1.45 and 4.67 ±0.99 mmoL/mL,
respectively, which were lower than those in the control group (8.51 ±
1.22 and 7.66 ±1.06 mmoL/mL); while the test group’s total swimming
time was signicantly prolonged to 47.95 ±7.16 min comparing with
the control group (39.42 ±5.11 min). The previous results illustrated
that the AbEXP1-a, an intracellular polysaccharide of AbM, had a
powerful antifatigue effect. Additionally, the molecular weight of
AbEXP1-a was within the range of macromolecular soluble active
polysaccharides, which indicated that AbMP could effectively improve
immunity. Chen et al. (2001) supplemented that AbM can produce a
strong stress effect on the circulatory system in Figure 3. AbMP can
improve the body’s ability to use stress factors, then it can effectively
increase the reexes of the "hypothalamic-pituitary-adrenal cortex
(HPA)" system in HPA-Axis and the body can quickly adapt to the harsh
environment through continuous self-regulation (Duan et al., 2003).
Therefore, they inferred that antifatigue effects of AbMP may correlate
Fig. 2. The experiment design of immunoregulation and antifatigue in vivo and test contents. The images were extracted from the BioRender app.
Fig. 3. The AbMP stress function promotion diagram in HPA-Axis. The images were extracted from the BioRender app.
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2194
with the adjustment of HPA system.
Previous experiments have revealed the antifatigue and immunor-
egulative function of AbMP in vivo test and raised the underlying
mechanisms by HPA-Axis system. However, AbMP’s absorption and
metabolic efciency in the human body still lack more clinical trials and
further discovery is in demand. Besides, promoting the deep fermenta-
tion of AbM mycelium technology may benet to developing functional
foods due to its high production efciency of crude polysaccharides.
4.2. Antidiabetic and anti-hyperglycemic effects
Type 2 diabetes is a group of complex heterogeneous metabolic
diseases affecting a large population in worldwide. Current theories for
the causes type 2 diabetes were attributed to the insulin-mediated
deciency of glucose uptake in muscle; impaired insulin action in the
liver and disruption of adipocyte secretion, etc.
α
-Glucosidase is an
enzyme that plays a crucial part in the nal step of the digestion process,
breaking down complex carbohydrates such as starch and glycogen into
monomers. Meanwhile, it is worth noting that
α
-glucosidase could
signicantly increase postprandial glucose in patients with type 2 dia-
betes, and the abnormal value of the
α
-glucosidase enzyme could also
lead to organ or tissue dysfunction, which in turn causes the develop-
ment of type 2 diabetes (Lin and Sun, 2009; Wei et al., 2019). However,
long-term consumption of existing antidiabetic drugs is supposed to
occur multiple undesirable problems, for instance, side effects of insulin
and oral hypoglycemic drugs (Etxeberria et al., 2012; Kim et al., 2005).
Thus, Teng et al. (2017) claried that there is a growing demand for
natural products which can potentially inhibit
α
-glucosidase because of
its role as the target of various drugs and inhibitors that regulate glucose
metabolism in the human body. Fortunately, edible mushrooms may
potentially prevent the secretion of
α
-glucosidase (Su et al., 2013;
Stojkovic et al., 2019). Wei et al. (2019) demonstrated AbM extracts’
inhibition of
α
-glucosidase activity through the method designed by
Chen and Kang (2014). 10–200
μ
g/mL AbM ethanol extracts (EE) and
ethyl acetate extracts (EA) were placed in the 96-well microplate, fol-
lowed by adding 0.1 U/mL
α
-glucosidase in sodium phosphate buffer.
And then the absorbance value at 405 nm was measured by ultraviolet
spectrometry, and the inhibition rate was calculated. It can be observed
that the AbM EE and EA at 8 mg/mL exhibited 64.86% and 73.45%
α
-glucosidase inhibition capacity in vitro. Therefore, it can be hypothe-
sized that AbM exhibited inhibition of the
α
-glucosidase under the action
of organic extracts like EE and EA, which also offered potential use for
anti-diabetic drugs. But Wei et al. (2019) ndings still lacking data from
in vivo and clinical trials and warrant further investigation, such as the
inhibitory effects can be achieved in vivo, and whether the concentra-
tions of the EE and EA extracts can be replicated in humans. And more
clinical data may give noteworthy assistance to medical practitioners in
the development of novel anti-diabetic drugs.
In addition, hyperglycemia may affect people with diabetes, which
would induce severe complications that require emergency care, such as
a diabetic coma ("Hyperglycemia in diabetes - Symptoms and causes",
2022). Kim et al. (2005) applied chromatography with Sephadex G-50
column to purify the AbM extracted β-glucans. And oligosaccharides
(AO) were derived from hydrolyzing β-glucans with an endo-β-(1 →
6)-glucanase from Bacillus megaterium. The body weight and
anti-hyperglycemia effects of β-glucans and AO were determined in
diabetes rats, respectively. The diabetes rats were induced by 55 mg/kg
streptozotocin through intraperitoneal administration for six weeks in
vivo. The rats were divided into four groups; normal control; diabetic
control; β-glucans treated, and AO treated. The body weight of rats in
each group decreased compared with the control group, which met the
agreement of Prince et al. (1998) research. The nal body weight of rats
treated with β-glucans and AO signicantly decreased from 268.8 g to
252.6 g and 263.2 g, respectively (p <0.05). Moreover,
anti-hyperlipemic indicators: serum triglyceride and serum total
cholesterol prominently decreased in β-glucans, and AO-treated groups
compared with the control group (p <0.05). Moreover, the hypergly-
cemia may also be inhibited by AbMP, as evidenced by Duan and Ma
(2005). An alloxan-induced mice diabetes model was established, and
concentrations of 4% and 8% AbMP were treated; the blood sugar
content of mice with both AbMP concentrations in the experimental
group was signicantly lower than that of the diabetes model (p <0.05).
Thus, they claimed that a specic concentration of AbM polysaccharide
had a hypoglycemic effect. On the other hand, Kim et al. (2005) studies
also indicated that β-glucans and AO inuenced insulin secretion; both
β-glucans and AO treated rats’ insulin concentration in pancreatic islets
elevated to 3.79 ng/mL and 4.89 ng/mL from 0.21 ng/mL in the diabetic
control group.
AbM exhibited
α
-glucosidase suppression activity in vitro, effectively
inhibiting type 2 diabetes. Additionally, the hyperglycemia in those with
diabetes could be veried by certain concentration of AbMP, β-glucans
and its derivation, AO may have glucose-lowering activity by stimu-
lating insulin secretion in the pancreatic islets in vivo, which may be an
essential source of latent functional food applications. However, the
antidiabetic activity and effectiveness of AO in developing alternative
drugs were twice superior to that of β-glucans (Kim et al. 2005). It can be
speculated that the monosaccharide hydrolyzed from glucans may have
a better antidiabetic effect than β-glucans and correlated anti-diabetes
activities tests for different animal models and clinical trials should be
conducted. In addition, during the production of AbM-based functional
foods, suitable ratio of monosaccharides to β-glucans content may be
more effective than aqueous solutions or AbMP extracts, but more in vivo
and in vitro experiments need to be veried. Moreover, the underlying
hypoglycemia mechanisms of cell models is quite limited, which still
need further experiments.
4.3. Hepatoprotective activity
Liver diseases have become one of the leading causes of mortality
worldwide in recent years (Dinakar et al., 2010). Traditionally, allo-
pathic medical practice has no satisfactory for severe liver disease
because the absent of reliable hepatoprotective drugs (Singh et al.,
2003). Although several potential drugs have been identied, herbal as
natural medicines were considered effective and safe alternatives to
treat liver diseases, considering the side effects of modern medicines,
doubtful efcacy, and safety. Thus, AbM as an edible medical fungus
aroused researchers’ interest for its potential hepatoprotective activity
(Madhu Kiran et al., 2012; Muthulingam et al., 2016). In general,
elevated levels of serum marker enzymes are generally considered to be
one of the most sensitive indicators of liver injury (Kapil et al., 1995).
Increasing levels of alanine aminotransferase (ALT) and aspartate
aminotransferase (AST) are indicators of cellular leakage and loss of
functional integrity of cell membranes in the liver (Drotman and Law-
horn, 1978). Furthermore, alkaline phosphatase (ALP), the
membrane-bound enzyme, is the prototype of mentioned enzymes that
reects pathological changes in bile ow (Ploa and Hewitt, 1989).
Muthulingam et al. (2016) applied the carbon tetrachloride (CCl
4
)
model to discovered hepatic damage recovery by AbM extracts as
illustrated in Figure 4. Mice were orally administrated with 250 and 500
mg/kg BW AbM aqueous extracts, and 1 ml/kg BW CCl
4
was given at the
same time for seven days. Initially, the activities of ALT, AST and ALP
signicantly improved in mice’s livers combating the CCl
4
induced
damage. However, serum elevated AST, ALT, ALP, bilirubin, and
cholesterol activities signicantly decreased, with an increase of serum
protein levels after 28 days of AbM extracts continuous treatment (p <
0.05). Serum AST and ALT levels were reduced to normal levels, indi-
cating the plasma membrane stabilization, which was consistent with
the accepted view that serum transaminase levels return to normal with
liver parenchymal healing and liver cell regeneration (Thabrew et al.,
1987). Therefore, they claimed that AbM aqueous extracts can poten-
tially inhibit the increase of ALP activity while reducing the elevated
bilirubin level, suggesting that it may stabilize mice’s liver function
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2195
during CCl
4
-induced liver injury. Furthermore, Zhang et al. (2016)
recognized the protective effect of AbMP liquid fermentation medium on
alcoholic liver injury in mice. They found that when the extracellular
polysaccharide content of AbM was 3.58 ±0.13 g/L, the levels of ALT
and AST in the test group were signicantly decreased (p <0.05), sug-
gesting that the AbMP fermentation solution also had protective effect
on alcoholic liver injury in mice. In a word, the aqueous extract of AbM
and AbMP was evidenced its hepato-therapeutic effect in vivo, and it may
be an alternative to existing drugs for the liver, but the effectiveness on
the human liver is still unclear, and toxicological testing also needs to be
rened to determine if functional food applications are possible. How-
ever, as a natural edible mushroom, AbM undoubtedly has broad
application prospects for replacing medicines or health supplements for
fewer side effects and a lower price.
4.4. Antitumor activity
There is currently little direct evidence for its specic anticancer
substances and mechanisms (Fujimiya et al., 1998). Ergosterol was rst
isolated through gel column chromatography and claimed to be an
antitumor compound in the AbM lipid fraction (Takaku et al., 2001).
Sarcoma 180 – bearing mice were orally administrated ergosterol at
doses of 400 and 800 mg/kg for 20 days, and which showed prominently
inhibition of tumor growth without side effects induced by other cancer
chemotherapy drugs such as thymus and spleen weights decreased.
Additionally, they intraperitoneally administrated 10, 50, 100 and 200
mg/kg BW ergosterol to the mice, and the results indicated that ergos-
terol suppression ratio could reach 20.6 %, 57.1 %, 65.8 % and 84.7 %,
respectively. Therefore, ergosterol or its metabolites were speculated to
might be involved in inhibiting the formation of tumor neo-
vascularization. Further research may focus on the possible synergistic
effect by other bioactive compounds in AbM and determine the under-
lying mechanism of ergosterol action at the cellular level. On the other
hand, a polysaccharide-protein complex named antitumor organic sub-
stance Mie (ATOM) was cultivated from AbM mycelia by Ito et al. (1997)
and they veried in vivo. The ATOM highly exhibited its antitumor ac-
tivity efciency on the model of Sarcoma 180 in mice which were
implanted ATOM subcutaneously at 10 and 20 mg/kg/day BW and
detected its ability against Ehrlich ascites carcinoma, Shionogi carci-
noma 42 and Meth A brosarcoma at 50 and 100 mg/kg/day. Moreover,
the ATOM exhibited no direct cytotoxic action on tumor cells in vitro;
thus, the authors indicated that the tumor growth inhibition was
attributed to immunological host-mediated mechanisms. But the ex-
periments for this polysaccharide-protein complex still remains at the
level of animal models, more experimental data from human beings
would be valid for deeper applications such as adjunct to an antitumor
drug or the supplement to a chemotherapy drug. Apart from that,
Mizuno et al. (1990) demonstrated that several polysaccharides,
including FI
0
-a-β (Mw about 50,000), FA-1-a-⍺ (Mw about 2,000,000),
FA-a-β and FA-2-b-β extracted from AbM which revealed a noticeable
antitumor ability. They applied ion-exchange chromatography and gel
ltration to purify and then isolate the polysaccharides and evidenced
their antitumor activity using Sarcoma 180/ICR-JCL mice for their
cytotoxicity in vivo. The control group mice had a tumor size of about 26
cm
3
at the third week, and after intraperitoneally 10 mg/kg FI
0
-a-β,
FA-1-a-⍺, FA-a-β and FA-2-b-β injected treatment for ten days, the tumor
size had decreased to 7.88 cm
3
, 1.80 cm
3
, 0.87 cm
3
, 1.23 cm
3
with
70.51, 93.32, 96.77 and 95.44 inhibition ratio, respectively.
Currently, most known compounds with tumor-killing activity and
their synthetic derivatives had insufcient tumor selectivity; chemo-
therapy regimens only responded to a few tumor diseases, such as he-
matopoietic malignancies. However, Fujimiya et al. (1998) claimed that
AbM inhibitory effect can enhance the host’s immunity against tumor
growth. The authors separated acid-treated fraction (ATF) from AbMP
and performed antitumor experiments on BaLB/c mice. By nuclear
magnetic resonance (NMR) analysis, the main components of ATF with
the highest anticancer activity were (1 → 4)-⍺-D-glucan and its branch
(1 → 6)-β. Then they set up a model with syngeneic MethA tumor cells
subcutaneously injected; the right ank was named primary tumor, and
the left was metastatic one. The distant primary tumor in the right blank
could be observed that was inltrated by natural killer cells in vivo.
Electrophoresis and DNA fragment assay could also clarify that
apoptotic processing was exhibited in tumor cells by ATF induced
directly in vitro. Moreover, ow cytometry analysis also exhibited that
ATF could increase the expression of the antigen Apo2.7 on the mito-
chondrial membrane of tumor cells, while it did not affect normal mice
and IL-2-treated splenic monocytes, thus, which could be conrmed
selective toxicity of ATF polysaccharide to tumor cells.
On the other hand, Murakawa et al. (2007) comprehensively found
the AbM water extracts’ antitumor activity on myeloma cells with the
combination of marine phospholipid liposomes. They designed experi-
mental group as below; (1) control; (2) 1.0 mg/mL squid phospholipid
liposome alone; (3) 0.5 mg/mL AbM water extract alone; (4) 1.0 mg/mL
squid phospholipid liposome with 0.5 mg/mL AbM water extract in
simple mixture; and (5) 1.0 mg/mL squid phospholipid liposome with
0.5 mg/mL AbM water extract partially encapsulated. The schematic
diagram of group 5 is illustrated in Figure 5 below. The BALB/c nu/nu
mice were inoculated subcutaneously with Sp-2 myeloma cells, and
tumor sizes and weight were measured after 20 days of implantation.
Previous studies illustrated that β-(1, 6)-glucan with β-(1, 3)-branched
chains exhibited strong anticancer ability, which would have a promo-
tion immunological competence when cooperated with liposomes
(Kodama et al., 2002). The experiment results exhibited that β-glucan
had a signicant inhibition effect on the growth of myeloma sp2 tumor
Fig. 4. The experimental process of CCl4-induced liver injury with corresponding target testing items. The images were extracted from the BioRender app.
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2196
cells in vivo (p <0.01), and the tumor disappeared in the partially
encapsulated form administrated group (group 5). According to
knowledge from Fujimiya et al. (1998) and Takaku et al. (2001), the
mechanisms behind the suppression of the tumor cells may be attributed
to AbM combined with liposomes cytotoxicity to tumor cells, interfer-
ence with tumor angiogenesis. In addition, we could suggest that AbM
constituents extract encapsulated with marine phospholipid liposomes
could modulate tumorigenesis and might be useful for myeloma sp2
disease treatment based on Murakawa et al., (2007) report.
Generally, a few AbM extracts were demonstrated and identied for
their antitumor activity in mentioned studies, but some unclear mech-
anisms remain partially understood. Polysaccharides such as FI
0
-a-β, FA-
1-a-⍺, FA-a-β and FA-2-b-β, isolated from AbM aqueous which revealed
antitumor activity and was responsible for immunological enhance-
ment. On the other hand, (1 → 4)-⍺-D-glucan in ATF extracted from AbM
had a selective antitumor activity that could induce apoptotic processing
in tumor cells. Compared with other antitumor drugs, ATOM and
ergosterol extracted from AbM both evidenced no cytotoxic effect on
180 sarcoma cells in vitro. More clinical trials may be conducted to verify
its effectiveness on the human body, and then it could be used as a
natural substitute for existing antitumor drugs. Moreover, the AbM
water extracts encapsulated in marine phospholipid liposomes can
regulate the occurrence of myeloma sp2 disease, which may be poten-
tially helpful for treating myeloma disease in the future. In the future,
more kinds of marine organisms not only the liposome binding of squid
can be used to encapsulate AbM for antitumor experiments. These
combinations may exist enormous commercial value for drug develop-
ment and the emergence of novel functional food.
4.5. Anti-inammatory effect
Inammation is involved in the primary pathological process of
many diseases, and which is an overreaction of the immune system. This
may attribute to stimulation of xenobiotics, oxidative stress, or patho-
logical processes such as injury and infection that are closely related to
the occurrence, development, and cure of many diseases (Lawrence
2009; Zhao et al., 2004). Appropriate stimulation of macrophages by
immune-related cytokines can protect the host from various pathologies
and cancers, but excessive accumulation of pro-inammatory cytokines
and free radicals from macrophages can also lead to severe inamma-
tory diseases, such as asthma, inammatory bowel disease (IBD) and
rheumatoid arthritis (Medzhitov, 2008; Pahwa et al., 2022). In the past,
anti-inammatory drugs were broadly divided into two groups: steroids
and non-steroids. In recent years, more and more attention has been paid
to the anti-inammatory effects of some plants and synthetic drugs with
sound anti-inammatory effects and mild adverse reactions (Zhao et al.,
2004).
Zhao et al. (2004) suggested AbMP had anti-inammatory effects on
acute, subacute, and immune-epidemic inammation in vivo. Thus, the
authors established four mouse models: ear inammation induced by
paraxylene, foot swelling induced by carrageenan, cotton ball granu-
loma and adjuvant arthritis in rats. They injected them peritoneally with
40, 80 and 160 mg/kg AbMP to study their anti-inammatory effects. As
a type of rheumatoid arthritis, the primary lesion of adjuvant arthritis in
rats was mainly a local acute inammatory reaction. In their experi-
ment, joint swelling could reach the peak at 18 h after inammation and
gradually decrease after three days. The results indicated that different
doses of the AbMP group had a signicant inhibitory effect on adjuvant
arthritis in rats compared with the normal saline group after 18 h and
three days based on the swelling rate of the inammatory site (p <0.05),
and which had no signicant difference with the indomethacin group.
However, the mechanism of action remained unclear. Similar results
could be obtained when acute inammation in mice induced dimethyl
benzene and subacute inammation of tampon granuloma.
As previously described, water-soluble AbMP (Mw 2.058 ×10
3
ku)
was extracted from AbM and puried using DEAE—sepharose fast ow
chromatography column by Liu et al. (2017) and was generally
acknowledged that have remarkable anti-inammatory activities. As the
main inammatory factor, TNF-
α
can promote T cells to produce various
inammatory factors to promote inammatory response, and nitric
oxide (NO) would regulate macrophages to phagocytose pathogenic
microorganisms and destroy pathogenic biological macromolecules.
Although previous studies indicated that AbMP could stimulate the
secretion of tumor necrosis factor TNF-
α
, IL-6, IL-10 and IFN-γ (Fu et al.,
2015; Yang et al., 2013), while lipopolysaccharides (LPS) would
over-stimulate macrophages when excessive (LPS) accumulated in the
body and then release inammatory mediators such as TNF-
α
, NO, and
ILs, thereby triggering the body’s inammatory response (Zhao et al.,
2010). In order to verify the anti-inammatory activity and mechanism
of AbMP, Liu et al. (2017) have studied the secretion of TNF and NO in
lipopolysaccharide (LPS)-induced inammation. TNF-
α
and NO secre-
tion were measured when adding 62.5, 125, 250, 500 and 1000
μ
g/mL
AbMP to 10 and 50
μ
g/mL lipopolysaccharide-induced inammation
mice model. The experimental results validated that AbMP signicantly
Fig. 5. The squid phospholipid liposome combined with partially encapsulated AbM extracts for myeloma sp2 tumor suppression. The images were extracted from
the BioRender app.
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2197
inhibited the secretion of these two cytokines, TNF-
α
and NO, in a
concentration-dependent manner (p <0.05), suggesting its
anti-inammatory activity. Specically, the TNF-
α
secretion revealed a
dose-response relationship at a relatively high concentration (250, 500
and 1000
μ
g/mL AbMP), and the NO concentration also signicantly
decreased when AbMP was lower than 250
μ
g/mL. However, the un-
derlying mechanisms behind it were still unclear; the authors proposed
it may be due to the suppression of phagocytosis and activation of
macrophages by AbMP. As summarized below, AbMP extracted could
potentially inhibit the production of inammatory cytokines from
macrophages which were induced by LPS (Figure 6). Another explana-
tion would be the anti-inammatory effects of down-regulated mRNA
and inducible Nitric Oxide Synthase (iNOS) expression levels of TNF-
α
,
but both hypotheses needed further addressed.
AbMP also exhibited anti-inammatory activities in current clinical
studies. AndoSan™ was conrmed as an anti-inammatory medicine
composed of AbMP, Hericium erinaceus (He) (14.7%) (LEE et al., 2000)
and Grifola frondosa (Gf) (2.9%) (Adachi et al., 1994) can treat CD by
reducing pro-inammatory cytokines in patients (Therkelsen et al.,
2016). Therkelsen et al. (2016) conducted a 21-day trial in 50 symp-
tomatic CD patients randomly assigned to receive daily oral AndoSan™
or placebo. The AndoSan™ group had signicantly improved scores for
symptoms, fatigue, and health-related quality of life, while the placebo
group had no signicant changes (p<0.001). Therefore, they assessed
that CD patients with mild to moderate symptoms might benet from
AndoSan™ as a safe supplement to conventional medication.
So far, AbMP has been demonstrated its potential medicinal value for
various anti-inammatory effects on acute, subacute, and immune-
epidemic inammation in vivo, and other diseases such as adjuvant
arthritis, otitis, and foot swelling may exist in humans. The follow-up
researchers should focus on establishing human inammation models
and exploring their anti-inammatory effects on the body. Moreover,
AbMP has exhibited its effectiveness in treating CD patients with mild or
moderate symptoms. Natural alternative medicine that containing
AbMP could be considered as a good choice for CD patients.
4.6. Antioxidant effect of AbM
It is well-known that the oxidation process is a fundamental reaction
in living organisms while continuous production of reactive oxygen
species (ROS) in vivo can lead to tissue damage and apoptosis. Human
ageing and certain diseases, including diabetes, cancer and atheroscle-
rosis, have been conrmed to be closely related to ROS (Calabrese et al.,
2006; Eastwood, 1999; Halliwell and Gutteridge 1999; Morton et al.,
2000). Thus, antioxidants aroused people’s interest in their potential
organisms protected from ROS, such as superoxide anion radical (O
2
•−
)
and hydrogen peroxide (H
2
O
2
). These by-products of normal meta-
bolism are produced during respiration, hydrolysis, etc., and which can
attack biomolecules such as lipids, proteins, DNA, and RNA, resulting in
cellular or tissue damage associated with degenerative diseases (Jung
et al., 1999). AbM is a natural food consisting of abundant poly-
saccharides, polyphenols and avonoids that were considered an anti-
oxidant with scavenging free radicals’ ability by scientists (Vellosa et al.,
2006; Hakime-silva et al., 2013).
In recent studies, Wu et al. (2014) comprehensively evaluated the
efciency of extraction ways to obtain three natural polysaccharides
(AbMP-F, AbMP-V, AbMP-A) antioxidant activity from AbM. AbMP-F,
AbMP-V, and AbMP-A are dened as freeze drying, vacuum drying
and air drying, respectively. The polysaccharides accounted for 12.28%,
11.54% and 11.19% of AbMP-F, AbMP-V and AbMP-A samples,
respectively. Moreover, their uronic acid followed the same order for
28.19%, 25.72% and 24.87% for AbMP-F, AbMP-V and AbMP-A,
respectively. The polysaccharides isolated from AbMP-F exhibited sig-
nicant differences in their content (p <0.05), suggesting freeze drying
was a better way of AbMP extraction. The antioxidant activity was
conducted for testing their radical scavenging ability, and four assays
Fig. 6. The potential mechanism of inhibition of inammatory cytokines by AbMP. The images were extracted from the BioRender app.
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2198
including hydroxyl radical, DPPH radical, ABTS free radical scavenging
ability and Fe
2+
-chelating ability assay with ascending AbMP concen-
tration. Ascending concentrations from 300, 600, to 900
μ
g/mL, all
showed positive concentration-dependent to their corresponding anti-
oxidant activities in all extracts for each assay. Specically, at 300
μ
g/mL, all three types of AbMP exhibited more than 50% scavenging
ability of ABTS free radical. Hydroxyl radical scavenging activity of both
AbMP-F and AbMP-V exhibited relatively high levels at all concentra-
tions, but the scavenging ability of AbMP-A to hydroxyl radicals was
always lower than 50% in the concentration range of 0.1–1.0 mg/mL.
But their chelating ability on Fe
2+
, scavenging activity on DPPH radi-
cals, and two previous assays showed a decreasing order of
AbMP-F>AbMP-V>AbMP-A. AbMP-F illustrated its higher antioxidant
abilities on scavenging hydroxyl radical, DPPH free radical, ABTS free
radicals, and ferrous ion chelating ability for its relatively highest
polysaccharides yield and uronic acid content. The author also sug-
gested that higher uronic acid content may potentially correlate with
AbMP’s antioxidant activities. Hence, researchers could devote to
discover the relationship of uronic acid with antioxidant ability. Addi-
tionally, the synergistic effect of uronic acid and AbMP alsoworth to be
determined.
Hakime-silva et al. (2013) obtained AbM aqueous macerated extracts
and they evaluated its ability against oxidative stress through various
antioxidant assays including (1) inhibition of oxidative enzymatic pro-
cess and cellular oxidative stress and (2) direct action over-reactive
species. The oxidative enzymatic process of HRP and MPO was con-
ducted by chemiluminescence with 100% suppression. Furthermore, the
ability to scavenge ROS, HOCl and superoxide anion radical O
2
• −
were
evaluated and exhibited 62% and 87% inhibition rate in vitro, respec-
tively. The authors also illustrated that AbM aqueous macerated extracts
could reduce ROS levels by inhibiting various reactive species or inter-
fering with enzymatic ROS generators. As for cellular oxidative stress,
the oxidative burst of polymorphonuclear neutrophils (PMNs) was
suppressed by 80% in their experiment. Another assessment of AbM
antioxidant activities was evaluated by Wei et al. (2019). AbM ethanol
extracts (EE) and ethyl acetate extracts (EA) were analyzed by DPPH,
ABTS radical scavenging, reducing power using K
3
Fe(CN)
6
in vitro. In the
DPPH study, the EE and EA exhibited 54.91% and 56.01% scavenging
ability at 500
μ
g/mL, with no signicant difference (p >0.05).
Furthermore, EE and EA in the present study contained 23 and 36 mg/g
of total phenolic and organic content from AbM extracts, respectively,
which mainly contributed to their antioxidant ability (Carvajal et al.,
2011). However, the ABTS assays suggested EA had better scavenging
activity than EE with lower IC
50
. The concentration of EE to inhibit 50%
of ABTS was 304
μ
g/ml; in contrast, 161
μ
g/ml of EA achieved the same
ratio. As for the reducing power of K
3
Fe(CN)
6
, the results illustrated a
concentrated-dependent in AbM extracts. The author applied gradient
concentration (156, 313, 650, and 1,250
μ
g/ml EE and EA), and the
reducing activity showed 0.24, 0.44, 0.94, and 1.45 for EE and 0.28,
0.50, 0.95, and 1.29 for EA, respectively.
AbM also exhibited tissue peroxidative damage protection and in-
hibition of abnormal antioxidant levels in CCl
4
-induced hepatotoxicity
in male albino rats. According to the previous study by Muthulingam
et al. (2016), CCl
4
can be used to induce liver tissue damage in rats. The
control experiment was adopted as follows: (1) control group (no
treatment), (2) only AbM extracts treatment and (3) only CCl
4
treat-
ment, (4) AbM extracts then treated with CCl
4
and (5) treatment with
CCl
4
followed by AbM extracts treatment. Various enzymes in serum,
such as ALT, AST and non-enzyme antioxidants glutathione, vitamin C,
and vitamin E were determined. Nonenzymatic antioxidants such as
reduced glutathione (GSH), vitamin C and vitamin E played an excellent
role in protecting cells from oxidative damage. GSH in the blood
maintains cellular levels of the active forms of vitamin C and vitamin E
by neutralizing free radicals (Aldridge, 1981; WINKLER, 1992).
Compared with the control group, the nonenzymatic antioxidant levels
in the experiment treated with AbM extracts were increased, while the
percentage of vitamin C, vitamin E, and GSH in rats decreased from
100% (control group) to 65.7%, 46.8% and 60.2% in the CCl
4
treated
group after AbM extracts adding, respectively, which veried that the
AbM extract was involved in inhibiting oxidative damage. In addition,
when GSH levels were reduced, so did the levels of vitamin C in the cells,
suggesting that GSH, vitamin C, and vitamin E are closely related to each
other, which also indicated that the AbM extract got involved in the
process of inhibiting oxidative damage (Al-Dbass, Al- Daihan & Bhat,
2012).
In conclusion, AbM is a natural source of antioxidant compounds
such as polysaccharides and phenolic compounds, and all of them have
shown decent antioxidant activities in vivo and in vitro. Table 2 sum-
marizes the antioxidant constituents with their activity test in previous
studies. Moreover, the freeze-drying method is a better way to prepare
AbMP, which can extract more polysaccharides, neutral sugar, and
uronic acid, which may achieve higher efciency on antioxidant sub-
stances production in the food industry.
4.7. Other health-promoting functions of AbM
In addition to the above-mentioned health functional improvement
of AbM, it also exhibited its antibacterial infection, antimutagenic and
anticarcinogenic effects on previous studies.
Table 2
The AbM extracts and antioxidant activities with corresponding assays.
Extract and
compounds
Methods Activities References
Polysaccharides
(AbMP-F,
AbMP-V, AbMP-
A)
DPPH and ABTS,
Hydroxyl radicals
scavenging; Fe2+-
chelating ability
Antioxidant
activities followed
decreasing order of
AbMP-F>AbMP-
V>AbMP-A; ABTS:
300
μ
g/mL for
more than 50%
scavenging ability
Wu et al.
(2014)
AbM aqueous
macerated
extracts
Enzymatic and
cellular oxidative
stress (HRP, MPO
and PMNs) and
direct action over
ROS (HOCl and
O2• − )
Enzymatic
oxidative stress of
HRP and MPO:
100% suppression;
Cellular oxidative
stress of PMNs:
80% inhibition;
HOCl and
superoxide anion
radical O2• − were
inhibited with 62%
and 87%
Hakime-silva
et al. (2013)
AbM ethanol
extract and ethyl
acetate extracts
Reducing power
(K3Fe(CN)6),
Hydroxyl, DPPH
and ABTS radicals
scavenging in vitro
Reducing power of
K3Fe(CN)6:
concentrated-
dependent
inhibition for EE
and EA; DPPH:
54.91% and
56.01% scavenging
ability at 500
μ
g/
mL EE and EA,
respectively; ABTS:
EE 304
μ
g/ml and
EA 161
μ
g/ml for
IC50
Wei et al.
(2019)
AbM aqueous
solution
Activities of non-
enzume
antioxidants:
glutathione,
vitamin C, and
vitamin E
Percentage of
vitamin C, vitamin
E, and GSH in rats
decreased from
100% (control
group) to 65.7%,
46.8% and 60.2%
in the CCl4 treated
followed by AbM
added group,
respectively.
Muthulingam
et al. (2016)
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2199
Food contamination and spoilage may lead to bacterial and viral
infections, thus, the development of food additives with antibacterial
activity to prevent food contamination has become necessary in the food
industry (Zhao et al., 2015). Bernardshaw et al. (2005) launched the rst
report on the anti-infective effect of AbM in vivo; 10
μ
L water extract of
AbM had reected its inhibitive effect of moderately virulent Strepto-
coccus pneumoniae serotype 6B activity on orally administered
NIH/OlaHsd mice. Although AbM extract lacked antibiotic effect against
pneumococcus in vitro, serum cytokine MIP-2 and TNF-
α
levels were
elevated in AbM extract-treated mice, suggesting that the protective
effect of AbM extracts was due to the involvement of the self-immune
system. The experiment served that AbM extracts may initiate an im-
mune response in animals and humans by boosting patients’ immunity.
According to the latest WHO estimates, cancer cases rose to 18.1
million, and the death toll could reach 9.6 million. Therefore, it is
imperative to adopt novel strategies to prevent from the cancer. Shen
et al. (2001) have studied the anticancer effects of two polysaccharides
(AbMP-II-
α
and AbMP-II-β) on human leukemia cells in vitro. The crude
polysaccharides of AbM were separated into ve components (I-V) by
DEAE-cellulose ion-exchange chromatography. Then the Sephadex
G-200 gel ltration column was used to isolate the components II into
AbMP-II-
α
and AbMP-II-β. 50, 100 and 200
μ
g/mL of AbMP-II-
α
and 5.8,
29 and 145
μ
g/mL AbMP-II-β were used to investigate the effect of
AbMP on the proliferation of leukemia cells. AbMP-II-
α
revealed a limit
inhibition effect at low concentrations. However, AbMP-II-β can inhibit
cell proliferation in a dose-wise fashion. On the contrary, the growth
cycle suppression of leukemia cells was signicant by AbMP-II-
α
. At 100
μ
g/mL, the inhibition rate could reach 77.8%-98.4%. The inhibition rate
could reach 25.51% after 72 h at a low concentration of 0.16
μ
g/mL.
Admittedly, the dosage and duration of AbMP-II-β were positively
correlated with its inhibition, reaching 100% after 48–72h treatment at
a high dose of 20–100
μ
g/mL. Bertollo et al. (2022) comprehensively
concluded that AbM had some therapeutic potential in terms of
anti-cancer effects as well as prevention and as adjuvant therapy to
chemotherapy and radiation therapy. The regulation of the immune
system is the key to its anti-cancer effect. Immunomodulation of AbM
interfered with the activity of cytochrome P450 enzymes to increase
chemotherapeutic function (Hashimoto et al., 2002). In addition, the
authors mentioned that the low molecular weight AbMP (LMW-AbMP)
in mushrooms could inhibit the activity of TNF-
α
and other cytokines
related to nuclear factor kappa B (NF-kB) activation, thereby reducing
the stimulation of NF-kB and E-activation of selectins. At present, the
mechanisms behind the anti-carcinogenic activity of AbM may not be
investigated enough in previous ndings which might need more clin-
ical trials to support their ndings.
On the other hand, several in vitro studies have indicated that AbM
extracts were involved in the inhibition of mutagenicity of benzo (a)
pyrene in the Ames Salmonella microsome assay (Osaki et al., 1994; Ito
et al., 1984). Delmanto et al. (2001) decided to investigate the
anti-mutagenic effect of AbM in vivo using the models of mouse bone
marrow micronucleus test (MNPCE) and peripheral blood micronucleus
test (MNRET). The results exhibited the antimutagenic effect of oral
gavage of 0.025 g/mL AbM extracts on 25 or 50 mg/kg cyclophospha-
mide (CP) intraperitoneal induced mice. It is worth noting that no single
AbM lineage was found to have anti-mutational activity in MNT tests. In
other words, the single lineage did not reduce CP-induced micronucleus
frequency in bone marrow or blood in mice compared to the mixed
lineage. The authors speculated that the antimutagenic component was
not evenly distributed across lineages and/or is not evenly present in
mushrooms at different times of the year. Thus, further investigation
should focus on specic bioactive compounds in AbM extracts that act
on the antimutagenic effect. Apart from that, the determination of a
single sequence with anti-mutation ability can be helpful for the sub-
sequent development of drugs and functional foods.
5. Impact of food processing
AbM as a functional food, is commercially cultivated in many
countries. However, few studies have mentioned about their impact in
food processing. In fact, different extraction methods of AbM composi-
tions would potentially affect its biochemical activity and lead to
numerous extraction efciencies during food commercial production.
The content of crude polysaccharides in the fermented mycelium of AbM
was much higher than AbM fruit body; thus, the research submerged
fermentation technology may greatly benet AbM in the future appli-
cation. Moreover, both boiling and microwave treatment of AbM
signicantly reduced the content of glucose, galactose, vitamin C and
mannose, but no signicant changes in their lipids content. Boiling and
microwaving signicantly reduced the species and relative content of
AbM phenolic compounds, resulting in reduced antioxidant capacity
such as DPPH scavenging activity (Sun et al., 2011). These effects need
to be taken into account when calculating food intake from cooked AbM.
On the other hand, Wu et al. (2014) suggested freeze-drying is a rela-
tively better way to extract AbMP with higher uronic acid content and
antioxidant effect than vacuum drying and air drying. Therefore,
selecting suitable extraction methods and parts plays a crucial role in the
study of the biological activity of AbM and maintain the nutritional
value of AbM as much as possible. Moreover, the impact during food
processing of several cooking methods such as steaming, frying and grid
splitting need further discoveries. At present, the related articles on AbM
food processing are quite limited; there are few studies on the effects of
different processing methods on the changes of nutrient components and
biochemical activity functions, and more comprehensive studies on the
effects of cooking methods on AbM can be conducted in the future.
5.1. Discussions and future application of AbM
AbM as a medicinal mushroom has showed several health-promoting
effects in vitro and in vivo. Polysaccharides were the widest extracts
isolated using heated aqueous or organic solvents such as ethanol and
ethyl acetate. According to previous studies, AbMP was believed to
affect its mechanism involved in the modulation of innate immunity.
More specically, AbM played a vital role in regulating macrophage
function. AbMP could regulate cytokines secreted by macrophages,
thereby reducing the production of inammation. Besides, the promi-
nent role of AbMP is to enhance and/or activate the macrophage im-
mune response, resulting in immune regulation, wound healing and
other therapeutic effects. These actions can be owned to the puried
β-glucan in AbM that can act on various cellular receptors related to the
initiation of immune responses. Nevertheless, there were no high-
quality clinical trial data available to evaluate the actual clinical ef-
cacy of puried β-glucan or compounds containing β-glucan (Chan et al.,
2009). Well-designed clinical trials can be conducted to verify the
effectiveness of β-glucans on human immunity and cancer cells in the
future. In conclusion, AbMP as a natural plant polysaccharide which
offers unique opportunities as a novel therapeutic adjuvant with bene-
cial immunomodulatory properties (Schepetkin and Quinn, 2006).
Apart from that, AbM extracts can be prepared without extraction,
exhibiting remarkable antidiabetic and antibacterial effects. Lipid and
polysaccharide-protein complex mainly showed antitumor effects in
vivo. These above-mentioned bioactive constituents with their target
functions were concluded in Table 3. And their future research di-
rections were also summarized in Figure 7.
According to Murakawa et al. (2007) research, encapsulating AbM
with marine organisms can contribute to a better anti-tumor activity,
thus, it can be speculated that the liposomes of the organisms may be
able to stimulate the activity of AbMP. Whether the combination of AbM
and liposomes can produce superior immunomodulatory function dur-
ing drug treatment is worthy of more in vivo data to validate. In addition,
AbM contains a variety of biologically active substances, and most ex-
periments currently utilized its aqueous solution of AbM for
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2200
experiments. On the other hand, many of the immunomodulatory sub-
stances may have synergistic effects on the target function. Therefore,
the purication and separation of the specic substances in AbM could
more effectively distinguish and identify biological activities and func-
tions in subsequent studies. The bioactivities of AbM have been based
mostly on in vitro and in vivo studies in mice. More clinical data and
epidemiological investigations are needed to evaluate immunomodula-
tion in humans at effective ingested doses of AbM as well as the chemical
identication and quantication of certain compounds. Moreover, the
content of heavy metals and the dose intake of mushroom alkaloids and
their derivatives are accompanied by potential toxicity and carcinoge-
nicity and that should be evaluated more comprehensively in AbM.
Despite the challenges, there is no doubt that AbM has advantages of
easy growth, low price, and rich nutrients which has great potential to
be utilized in the market as functional food and drug substitutes in the
future.
6. Conclusions
AbM is considered a medical mushroom for its immunological,
antitumor and anti-inammatory activity, owned to its various bioactive
compounds including polysaccharides, lipids, and phenols. On the other
hand, freeze-drying method was considered as the comparatively better
way to acquire AbMP with higher antioxidant effect. Meanwhile, deep
Table 3
The main bioactive compounds found in the AbM, doses and administration routes, and target functions.
Bioactive substances Components Dose and/or administration route Function References
Polysaccharide AbM polysaccharides (AbMP) 100, 150, and 200 mg/kg BW; oral Immunoregulation Li and Wei (2017)
Polysaccharide AbEXP1-a 0.2 mL/kg BW; ingest Anti-fatigue LI (2020)
Polysaccharide AbMP 100, 150, and 200 mg/kg BW; gavage Anti-fatigue Li and Wei (2017)
Polysaccharide β-Glucans 2% β-Glucans; oral Anti-diabetic Kim et al. (2005)
Polysaccharide β-Glucans 0.5 mg/mL AbM water extract; oral Antitumor Murakawa et al. (2007)
Polysaccharide AbMP 4% and 8% Hyperglycemia Duan and Ma (2005)
Polysaccharide AbMP (AbMP-F, AbMP-V,
AbMP-A)
300, 600, and 900
μ
g/mL (in vitro) Antioxidant Wu et al. (2014)
Polysaccharide Acid-treated fraction (ATF) 1 mg; injected intratumorally Antitumor Fujimiya et al. (1998)
Polysaccharide AbM polysaccharides (AbMP) 40, 80 and 160 mg/kg;intraperitoneal injection Anti-inammatory Zhao et al. (2004)
Polysaccharide AbMP-II-
α
and AbMP-II-β 50, 100 and 200
μ
g/mL of AbMP-II-
α
and 5.8, 29 and 145
μ
g/mL AbMP-II-β (in vitro)
Anticarcinogenic SHEN, SUN, LIU & GU (2001)
Polysaccharide FI
0
-a-β, FA-1-a-⍺, FA-a-β and
FA-2-b-β
10 mg/kg BW; Intraperitoneal injection Antitumor Mizuno et al. (1990)
Polysaccharide AbMP (Mw 058 ×103 ku) 62.5, 125, 250, 500 and 1000
μ
g/mL (in vitro) Anti-inammatory Liu et al. (2017)
Polysaccharide-protein
complex
Antitumor organic substance
Mie (ATOM)
10 and 20 mg/kg/day and 50 and 100 mg/kg/day;
subcutaneous
Antitumor Ito et al. (1997)
AbM extracts – 8 mg/mL EE and EA (in vitro) Antidiabetic Wei et al. (2019)
AbM extracts – 0.5 g/kg; oral Antioxidant Al-Dbass, Al- Daihan & Bhat,
(2012)
AbM extracts – 250 and 500 mg/kg BW; oral Hepatoprotective Muthulingam et al. (2016)
AbM extracts – 10
μ
L AbM aqueous; oral Antibacterial Bernardshaw et al. (2005)
AbM extracts – 0.025 g/mL; orally gavage Antimutagenic Delmanto et al. (2001)
AbM extracts AbMP 3.58 ±0.13 g/L;gavage Hepatoprotective ZHANG, LIN, DONG, WANG &
HAN, (2016)
AbM extracts – 100
μ
L AbM aqueous (in vitro) Antioxidant HAKIME-SILVA et al. (2013)
AbM extracts – 156, 313, 650, and 1,250
μ
g/ml EE and EA (in vitro) Antioxidant Wei et al. (2019)
Lipid Ergosterol 10, 50, 100 and 20 mg/kg BW; intraperitoneal 100, 200,
400 and 800 mg/kg BW; oral
Antitumor Takaku et al. (2001)
Fig. 7. The bioactive constituents with their target functions and future research directions of AbM. The images were generated by XMind app.
K. Huang et al.
Current Research in Food Science 5 (2022) 2190–2203
2201
fermentation of AbM mycelium and use of AbMP-derived AO may
potentially provide a better choice for functional foods such as diabetes.
Up to now, research on the immune regulation, diabetes, and antitumor
impact of AbM is gradually deepening, while polysaccharides such as
β-Glucans and ATF would be subjected to more epidemiological in-
vestigations. Taken together, most of the experiments are currently
concentrated in test tubes and animals, and there are relatively few
universal clinical trials for the effectiveness of human body. However, it
is believed that there would seek broad application prospects in the
future as natural therapeutic alternatives based on AbM extracts due to
their abundant nutritional value and health promotion activities.
Funding
This study is jointly supported by two research grants R202016 and
R202107 from BNU-HKBU United International College.
CRediT authorship contribution statement
Kaiyuan Huang: Investigation, Data curation, Formal analysis,
Software, Validation. Baojun Xu: Conceptualization, Methodology,
Software, Supervision, Funding acquisition, Project administration,
Validation, Writing – review & editing.
Declaration of competing interest
The authors declare that they have no known competing nancial
interests or personal relationships that could have appeared to inuence
the work reported in this paper.
Abbreviation
ABL. Hemagglutinin
AbM Agaricus blazei Murill
AbMP Agaricus blazei Murill polysaccharides
AIDS Acquired Immune Deciency Syndrome
ALP. Alkaline phosphatase
ALT. Alanine aminotransferase
AST Aspartate aminotransferase
ATF Acid-treated fraction
ATOM antitumor organic substance Mie
BW. Body weight
CCl
4
Carbon tetrachloride
CD Crohn’s disease
CP. Cyclophosphamide
DEAE Diethyl-aminoethanol
EA Acetate extracts
EA. Ethyl acetate extracts
EE. Ethanol extracts
EE. Ethanol extracts
Gf Grifola frondose
GSH. Glutathione
He Hericium erinaceus
HIV. Human immunodeciency virus
HRP. Horseradish peroxidase
HRP. Horseradish peroxidase
IL. Interleukin
iNOS Inducible Nitric Oxide Synthase
LPS. Lipopolysaccharides
MDA. Malondialdehyde
MNPCE Mouse bone marrow micronucleus test
MNRET Peripheral blood micronucleus test
MPO. Myeloperoxidase
MS Mass spectrometry
NF-kB Nuclear factor kappa B
NMR. Nuclear magnetic resonance
NO Nitric oxide
PMNs Polymorphonuclear neutrophils
ROS. Reactive oxygen species
SOD. Superoxide dismutase
TNF Tumor necrosis factor
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