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A Review of the Bioactive Compound and Medicinal Value of Cordyceps Militaris

Authors:
  • Northern Border University, Arar, Saudi Arabia

Abstract

For centuries, traditional Chinese medicine has relied on medicinal fungi as a panacea for strengthening the immune system and revitalizing the human body based on the belief that fungi have diverse biological functions. Modernly, Cordyceps militaris has also been a source of several bioactive components in pharmacology and medicine, such as cordycepin, ergosterol and polysaccharide. Cordyceps militaris has been the topic of numerous reviews; however, many of them have not focused on the bioactive compound and medicinal value of this fungus. In this mini-review, we compile recent data on, the latest molecular research, its bioactive compounds and medicinal value.
69
A Review of the Bioactive Compound and Medicinal Value of Cordyceps militaris
Abdulhakim Bawadekji*1, Khalil Al Ali 2, Mouhanad Al Ali 3
(Received 12/02/2016; accepted 26/03/2016)
Abstract: For centuries, traditional Chinese medicine has relied on medicinal fungi as a panacea for strengthening the
immune system and revitalizing the human body based on the belief that fungi have diverse biological functions. Modernly,
Cordyceps militaris has also been a source of several bioactive components in pharmacology and medicine, such as
cordycepin, ergosterol and polysaccharide. Cordyceps militaris has been the topic of numerous reviews; however, many of
them have not focused on the bioactive compound and medicinal value of this fungus. In this mini-review, we compile recent
data on, the latest molecular research, its bioactive compounds and medicinal value.
Keywords: Cordyceps militaris, Bioactive compound, Entomopathogenic fungus, Medicinal value.
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(Cordyceps militaris) 

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


   Cordyceps militaris           

    

Cordyceps militaris 
Cordyceps militaris
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Northern Border University
Journal of the North for Basic and Applied Sciences (JNBAS)
www.nbu.edu.sa
http://ejournal.nbu.edu.sa
 )

* Corresponding Author:
(1) Deanship of Scientific Research, Northern
Border University, P.O.Box 1321, Arar 91431,
Kingdom of Saudi Arabia.
DOI: 10.12816/0021378
e-mail: hakimbawadekji@gmail.com & bawadekji@nbu.edu.sa
 

A Review of the Bioactive Compound and Medicinal Value of Cordyceps militaris.
70
1. INTRODUCTION
For centuries, traditional Chinese medicine has
turned to medicinal fungi as a panacea for
strengthening the immune system and revitalizing
the human body, qualities that have usually been
attributed to their diverse biological activities, the
two most commonly used fungal species being
Cordyceps sinensis and Cordyceps militaris (Liu,
1994; Liang, 2007), of which
C. sinensis is more famous. Many active
compounds found in the two varieties have positive
effects not only on the human immune and
respiratory systems but in the treatment of several
diseases, such as renal, hyperglycemic and hepatic
dysfunctions (Song, Jeon, Yang, Ra, & Sung,
1998; Mizuno, 1999; Yun, Han, Lee, Ko, Lee, Ha,
& Kim, 2003; Choi, Par, Choi, Jun, & Park, 2004;
Yu, Wang, Zhang, Zhou, & Zhou, 2004). Although
C. sinensis and C. militaris have identical
medicinal properties, C. militaris is more easily
cultured (Zheng, Huang, Cao, Xie, & Han, 2011;
Dong, Lei, Ai, & Wang, 2012). It is a great source
of biomedical products (Ng & Wang, 2005) and a
more dependable fungus for extracting bioactive
components, such as cordycepin, ergosterol and
polysaccharides, that are used in pharmacology and
modern medicine (Das, Masuda, Hatashita,
Sakurai, & Sakakibara, 2010; Reis, Barros,
Calhelha, Ćirić, Van Griensven, Soković, &
Ferreira, 2013). Moreover, research has shown that
C. militaris also possesses the ability to infect and
parasitize lepidopteran insects, such as butterflies,
at different stages, infecting their pupae (or larvae),
living and developing there until finally killing and
mummifying them. Thereafter, C. militaris
produces fruiting bodies of 2 to 8 cm in length of
and a width of 0.5 cm (Han, Liu, Cao, & Chen,
2006; Hong, Kang, Kim, Nam, Lee, Choi, Kim,
Kim, Lee, & Humber, 2010).
C. militaris has been the topic of numerous
reviews, many of which have not focused on both
the bioactive compound and medicinal value
aspects of the fungus. Consequently, this mini
review will expand the bibliographic research into
those aspects to focus on the biological activities of
Coryceps militaris, and to reveal the importance of
this entomopathogenic fungus as valuable source
of medicinal compounds. In addition, to highlight
the need for further, molecular research to refine
the taxonomical position of Cordyceps spp. and the
necessity to better understand biochemical
synthetic pathway of its bioactive compounds.
2. TAXONOMY OF Cordyceps militaris
RELATED TO MOLECULAR
RESEARCH
C. militaris
(L.: Fr.)Link classified in the
subphylum Ascomycotina, Clavicipitales and
Clavicipitaceae. In previous studies, the
classification of this fungus based on
morphological characters, such as asci, ascospore
fragmentation, thickened ascus apices, part-spores,
and the arrangement of perithecia (Kobayasi, 1941,
1982; Mains, 1957, 1958). However, since that
system proved difficult to apply and inefficient in
differentiating between any two close species of
fungi, which the research attributed to various
factors, such as the environment and the effects of
diverse ecological habitat conditions on
morphological characters, there have been attempts
to offer alternate classifications using molecular
methods. For instance, Sung, Hywel-Jones, Sung,
Luangsa-ard, Shrestha, & Spatafora, (2007) used
molecular markers to refine the classification of
Clavicipitaceae that utilized seven loci: nuclear
ribosomal small subunits (nrSSU), nuclear
ribosomal large subunits (nrLSU), elongation
factor 1α (tef1), the largest subunits of RNA
polymerase II (rpb1), the second largest subunits
of RNA polymerase II (rpb2), β-tubulin (tub), and
mitochondrial ATP6 (atp6) (ibid). The results
indicated that most of the morphological characters
used in earlier classifications of Cordyceps spp.,
such as the arrangement of perithecia, ascospore
fragmentation, ornamentation, etc., were not
adequate to study phylogenetic relationships; they
did not provide phylogenetic information, whereas,
the pigmentation, texture and shape of the stromata
were principally phylogenetically informative
(ibid).
Journal of the North for Basic and Applied Sciences, Vol. 1, Number (1), Northern Border University, (2016 /1437H.)
71
3. BIOACTIVE COMPOUND AND
MEDICINAL VALUE OF Cordyceps
militaris
3.1. Cordycepin
In recent years, several bioactive compounds have
been extracted and examined. The first, and
primary, bio-active compound extracted was
cordycepin (3-deoxyadenosine), which is a
nucleoside analogue synthesized by C. militaris,
that possesses several pharmaceutical proprieties,
and is widely used in modern medicine (Zongqi,
2002; Tuli, Sandhu, & Sharma, 2014). It has also
been shown that cordycepin (3-deoxyadenosine)
displays antimicrobial, immunomodulatory and
anticancer effects (Ohta, Lee, Hayashi, Fujita,
Park, & Hayashi, 2007; Vitali, Petrelli,
Lambertucci, Prenna, Volpini, & Cristalli, 2012),
and that it is intracellularly transformed into its 5′
mono-, di- and triphosphates, which inhibits the
activities of several enzymes in the purine
biosynthetic pathway (Masuda, Urabe, Sakurai, &
Sakakibara, 2006).
3.2. Ergosterol peroxide
Several previous studies conducted to determine
the effect of ergosterol peroxide (CAS Number:
2061-64-5) extracted from C. militaris
demonstrated that ergosterol peroxide has a
significant activity against gastric cancer cell line
(Kim, Kim, Cai, Nam, Lee, An, Jeong, Yun, Sung,
Lee, & Hyun, 2001). Another also catalogued the
anti-inflammatory, anticancer effects of ergosterol
peroxide, that found that it suppresses
inflammatory responses through the inhibition of
the transcriptional activity of NF-kB and C/EBPb
and the phosphorylation of MAPKs. This study
also confirmed that ergosterol peroxide is one of
the most important antitumor sterols to be
produced by medicinal mushrooms -- although its
molecular mechanism still remains a mystery
(Kobori, Yoshida, Ohnishi-Kameyama, &
Shinmoto, 2007).
3.3. Adenosine
It is well documented in the literature that
adenosine is considered as the main nucleoside in
Cordyceps spp. (Yang, Li, Li, & Wang, 2007b).
Yang, Guan, & Li, (2007a) discovered a large
number of C. militaris adenosines although the
figure is low when compared to that discovered for
C. sinensis (Yang et al., 2007b). Other studies have
also documented that adenosine has an essential
role in the biochemical process and that it
possesses several pharmaceutical properties, such
as anti-inflammatory and anticonvulsant activities,
and that it can also be used to treat chronic heart
failure and, perhaps, even prevent tissue damage
(Ontyd and Schrader, 1984; Katakaze & Hori,
2000).
3.4. Fibrinolytic enzyme
A new bioactive compound similar to subtilisin-
like serine protease was extracted from Korean C.
militaris by Choi, Cha, Park, Kim, Lee, Park, &
Park, (2011), which was a fibrinolytic enzyme that
had a molecular mass of 34 kDa, with a sequence
alignment that indicated that the enzyme shared the
highest (68%) sequence identity with subtilisin
PR1J (Gene bank, CAC95048), that was isolated
from Metarhizium acridum. It is worthy of note
that the N-terminal sequence of the enzyme
contains 15 amino acids, different from those of
fibrinolytic enzymes found in other mushrooms.
Results obtained from research conducted to
investigate the effect of fibrinolytic enzyme
purified from C. militaris, also indicated that the
fibrinolytic enzyme has fibrin binding activity and
that it contributes to fibrin pathway degradation
(Kim, Sapkota, Park, Choi, Kim, Hiep, & Park,
2006; Choi, Par, Choi, Jun, & Park, 2011).
3.5. Xanthophylls
Recently, studies have reported that C. militaris
fruit-bodies contain essential bioactive
components. Research has also documented that
Cordyceps carotenoids are considered as some of
the most important active components present in C.
A Review of the Bioactive Compound and Medicinal Value of Cordyceps militaris.
72
militaris fruit-bodies (Dong, Wang, Ai, Yao, Sun,
Lei, & Wang, 2013).
Substantial research has also been conducted to
study the Cordyceps carotenoids that are isolated
from C. militaris fruit-bodies. For example, Dong
et al. (2013) identified and classified four new
xanthophylls: cordyxanthin-I, cordyxanthin-II,
cordyxanthin-III and cordyxanthin-IV, although
the chemical compositions and pharmaceutical
properties of these carotenoids are still to be
determined
3.6. Polysaccharides
Studies have reported that polysaccharide extracted
from the fruiting bodies of C. militaris has an
important medicinal value. Ohta et al. (2007)
isolated an acidic polysaccharide (APS) that
consisted of D-galactose (CAS number: 59-23-4),
L-arabinose (CAS number: 5328-37-0), L-
rhamnose (CAS number: 3615-41-6), D-xylose
(CAS number: 58-86-6) and D-galacturonic acid
(CAS number: 6556-12-3). In their study, Ohta et
al. (2007) examined the effect of acidic
polysaccharide (APS) on mice infected with the
influenza A virus. Interestingly, the results
demonstrated that when the polysaccharide was
administered, the virus titers decreased in the mice
and their survival rate increased. Moreover, the
study indicated that acidic polysaccharide (APS)
increased TNF-alpha and IFN-gamma levels in the
treated mice while also enhancing nitric oxide
(NO) production. Furthermore, the study also
noted that there was an induction of iNOS mRNA
and protein expressions in RAW 264.7 murine
macrophage cells. The study concluded that its
results proved that APS had anti-viral effects on
the influenza A virus through the modulation of the
immune function of macrophages (Ohta et al.,
2007).
In a subsequent, it was discovered that
polysaccharide (cordlan) extracted from C.
militaris has an antitumor activity (Kim, Kim,
Kang, Kim, Kim, Hong, Lee, Hong, Kim, & Han,
2010). The study examined the effects of
polysaccharide (cordlan) on the defect of dendritic
cells (DC), and established that polysaccharide
(cordlan) increased allogenic T cell stimulation but
that it decreased endocytosis. The study further
indicated that cordlan increased the
phosphorylation of the main signaling molecules
down-stream from TLR4. The study concluded that
its results demonstrated that cordlan induces DC
maturation via TLR4 signaling pathways (Kim et
al., 2010). In a corresponding study (Li, Li, Li,
Douz, & Gao, 2010), to examine the effects of
polysaccharides isolated from cultivated fruiting
bodies of C. militaris (CMP), it was observed that
CMP can inhibit mitochondrial injury and
mitochondrial swelling through scavenging
reactive oxygen species (ROS) and increasing anti-
oxidase activities, which the study concluded
indicated that CMP has both anti-aging activity and
pharmaceutical properties.
The investigation of Lin, Liu, Wu, Pang, Jia, Fan,
Jia, & Jia (2012) on the antioxidant effect of the
exopolysaccharide (EPS) isolated C. militaris SU5-
08, (which had been extracted from a strain of C.
militaris SU5), revealed that the EPS of C.
militaris SU5-08 had an antioxidant activity that
reinforced adaptive immune responses.
3.7. Antibacterial effect of Cordyceps militaris
Research recently undertaken to evaluate the
antibacterial activity of C. militaris against human
bacteria includes one in Korea to investigate the
antibacterial effect of the fungus on nine human
intestinal bacteria (Che, 2003). The results
indicated that the liquid culture of
C. militaris has growth-inhibiting activity against
several bacteria, such as Clostridium perfringens
and Clostridium paraputrificum, which the
research attributed to an active compound present
in the liquid culture of C. militaris, that the
research identified as cordycepin (3„-
deoxyadenosine). It is noteworthy from the study
that cordycepin did not reveal any adverse effects
against Bifidobacterium bifidum, Bifidobacterium
longum, Bifidobacterium breve, Bifidobacterium
adolescentis, Lactobacillus casei, or Lactobacillus
acidophilus. This outcome suggests that C.
militaris has at least one pharmacological action;
moreover, that cordycepin could be a valuable
antibacterial agent against various diseases caused
by Clostridium spp. (Ahn, Park, Lee, Shin, & Choi,
2000).
Journal of the North for Basic and Applied Sciences, Vol. 1, Number (1), Northern Border University, (2016 /1437H.)
73
3.8. Effect of Cordyceps militaris on fertility
C. militaris is prevalently used in traditional
medicine to improve sperm production and
enhance sexual activity; but no evidence to
corroborate the effects on humans. However, a
study on boars by Lin, Tsai, Chen, Hou, Hung, Li,
& Jeng (2007) conducted in Taiwan had surprising
results. Two groups of boars, one fed on a normal
diet (control group) and another on a C. militaris
mycelium supplemented diet (treated group).The
results revealed that sperm production was
considerably enhanced in the treated group;
moreover that sperm motility and morphology
were also significantly enhanced in the group.
3.9. Uses and health benefits of Cordyceps
militaris
C. militaris is widely used as food in Southeast
Asia, especially in China, Taiwan and Hong Kong
(Li, Guan, & Li, 2015). The fruiting bodies of C.
militaris are used to make traditional foods and
drinks, such as stewed chicken, duck, tea, and a lot
of other traditional staples (ibid). The fruiting
bodies are considered safe when consumed at less
than 2.5 g/kg of body weight. Li et al., 2015
examined the effects of cooking on the C. militaris
compounds, adenosine and cordycepin, and found
that the former compound decreased dramatically
when the fungus is steamed while the later
remained relatively stable.
Nowadays, the health benefits of the fruiting
bodies and mycelia of C. militaris are found in
pharmaceutical products and drugs. Cultures of C.
militaris have showed interesting properties, such
as utility in producing drugs for various diseases,
example, chronic bronchitis, kidney ailments and
pulmonary diseases, etc. (Dai, Fan, Wu, Xiao, &
Tian, 2007; Wang & Yang, 2006). Over 30
varieties of pharmaceutical products are now
available commercially (Huang, Lin, & Chen,
2010).
4. DISCUSSION
Some researchers have discovered that the
chemical components of the wild C. militaris and
the cultured C. militaris are similar (Tong, Kuang,
Wu, Zhang, & Ren, 1997; Jiang & Sun, 1999;
Wang, Lee, Chen, Yu, & Duh, 2012). For many
years, solid cultures were used to produce
industrial enzymes (Marques de Souza, Zilly, &
Peralta, 2002; Fenice, Giovannozzi, Federici, &
D‟Annibale, 2003) and nutrient enriched feeds
(Aguilar, Aguilera-Carbo, Robledo, Ventura,
Belmares, Martinez, Rodriguez-Herrara, &
Contreras, 2008; Vintila, Dragomirescu, Jurcoane,
Vintila, Caprita, & Maniu, 2009; Soltani, Al-Ali,
Othman, Malik, Elmarzugi, Aziz, & Al Enshasy,
2015). However, the process was found to be more
cost-effective than liquid and two-stage cultures.
At present, due to various reasons, such as host
specificity and scarcity, the fruiting bodies and
mycelia of wild C. militaris are expensive to
obtain. Consequently, current attempts to find and
develop alternative methods to extract its bioactive
components involve artificial cultivation, that
involves solid, liquid (including submerged and
surface liquid), and two-stage cultures (Junjun,
2007). Many researchers produce the fruit-body of
the fungus in vitro; some successfully grow C.
militaris and obtain fruiting bodies on brown rice
medium (Sung, Choi, Lee, Kim, Kim, & Sung,
1999; Sung, Choi, Shrestha, & Park, 2002). Other
scientists have also developed methods to
successfully grow fruiting bodies on insect pupae
(Harada, Akiyama, Yamamoto, & Shirota, 1995;
Sato & Shimazu, 2002). However, although solid
culture is cost-effective and appears to be a
promising culture technology, it takes considerable
time to yield fruiting bodies; the quality of the final
product is also not easy to manage, and solid
culture is not appropriate for large-scale industrial
production (Dong, 2013).
In recent years also, the liquid culture of
C. militaris has been widely studied to enhance the
production of valuable metabolites. This method
has many advantages, the main ones being those of
producing a high quantity of mycelia in a short
time and the reduction in the risk of contamination
(Hsieh, Tsai, & Shih, 2007; Kim, Hwang, Park,
Cho, Song, & Yun, 2002; Park, Kim, Hwang, &
Yun, 2004).
A Review of the Bioactive Compound and Medicinal Value of Cordyceps militaris.
74
A substantial amount of research has also been
conducted to shed light on the genetic and
molecular biology of C. militaris. Scientists have
identified several genes involved in the formation
of fruiting bodies, which is significant in
improving the production of bioactive compound
(Tuli et al., 2014). In order to enhance the
production of this valuable compound, scientists
have successfully obtained a C. militaris mutant
using different techniques, such as ion beam
irradiation (Das et al., 2010).
To our knowledge, there has been no report of any
toxicity in humans, and a recent study also
indicated that C. militaris can be used as a fresh
health food (Zhu, Pan, Yang, & Zhou, 2015).
5. CONCLUSION
As mentioned previously, the research paper study
has shown that the biological activities of the
Cordyceps militaris fungus are valuable sources of
medicinal compounds; however, further molecular
research is necessary to refine the taxonomical
position of Cordyceps spp. and in order to improve
the production of the medicinal compounds and
facilitate a better understanding of their
biochemical synthetic pathway.
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... Adenosine and cordycepin isolated from C. militaris exhibited several biological activities [34]. Adenosine has benefits for the prevention of ROS-related diseases such as tissue damage, is a therapeutic agent against chronic heart failure, and has anti-inflammation properties [31]. Cordycepin exhibited beneficial effects, such as broad-spectrum antibiotic activity, inhibition of cell proliferation, induction of cell apoptosis, as well as antioxidant activity [35,36]. ...
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Cordyceps is a rare naturally occurring entomopathogenic fungus usually found at high altitudes on the Himalayan plateau and a well-known medicinal mushroom in traditional Chinese medicine. Cordyceps contains various bioactive components, out of which, cordycepin is considered most vital, due to its utmost therapeutic as well as nutraceutical potential. Moreover, the structure similarity of cordycepin with adenosine makes it an important bioactive component, with difference of only hydroxyl group, lacking in the 3′ position of its ribose moiety. Cordycepin is known for various nutraceutical and therapeutic potential, such as anti-diabetic, anti-hyperlipidemia, antifungal, anti-inflammatory, immunomodulatory, antioxidant, anti-aging, anticancer, antiviral, hepato-protective, hypo-sexuality, cardiovascular diseases, antimalarial, anti-osteoporotic, antiarthritic, cosmeceutical etc. which makes it a most valuable medicinal mushroom for helping in maintaining good health. In this review, effort has been made to bring altogether the possible wide range of cordycepin’s nutraceutical potential along with its pharmacological actions and possible mechanism. Additionally, it also summarizes the details of cordycepin based nutraceuticals predominantly available in the market with expected global value. Moreover, this review will attract the attention of food scientists, nutritionists, pharmaceutical and food industries to improve the use of bioactive molecule cordycepin for nutraceutical purposes with commercialization to aid and promote healthy lifestyle, wellness and wellbeing.
... Antitumor, anti-diabetic, anti-inflammatory, antimicrobial, inhibit platelet aggregation, hypolipidemic, analgesic, immunomodulatory [15,[30][31][32] Adenosine ...
Article
Full-text available
Cordycepsis a rare naturally occurring entomopathogenic fungus usually found at high altitudes on the Himalayan plateau and a well-known medicinal mushroom in traditional Chinese medicine. Cordyceps contains various bioactive components, out of which, cordycepin is considered most vital, due to its utmost therapeutic as well as nutraceutical potential. Moreover, the structure similarity of cordycepin with adenosine makes it an important bioactive component, with difference of only hydroxyl group, lacking in the 30 position of its ribose moiety. Cordycepin is known for various nutraceutical and therapeutic potential, such as anti-diabetic, anti-hyperlipidemia, anti-fungal, anti-inflammatory, immunomodulatory, antioxidant, anti-aging, anticancer, antiviral, hepato-protective, hypo-sexuality, cardiovascular diseases, antimalarial, anti-osteoporotic, anti-arthritic, cosmeceutical etc. which makes it a most valuable medicinal mushroom for helping in maintaining good health. In this review, effort has been made to bring altogether the possible wide range of cordycepin’s nutraceutical potential along with its pharmacological actions and possible mechanism. Additionally, it also summarizes the details of cordycepin based nutraceuticals predominantly available in the market with expected global value. Moreover, this review will attract the attention of food scientists, nutritionists, pharmaceutical and food industries to improve the use of bioactive molecule cordycepin for nutraceutical purposes with commercialization to aid and promote healthy lifestyle, wellness and wellbeing
... Antitumor, anti-diabetic, anti-inflammatory, antimicrobial, inhibit platelet aggregation, hypolipidemic, analgesic, immunomodulatory [15,[30][31][32] Adenosine ...
Article
Full-text available
:Cordycepsisararenaturallyoccurringentomopathogenicfungususuallyfoundathighaltitudes on the Himalayan plateau and a well-known medicinal mushroom in traditional Chinese medicine. Cordyceps contains various bioactive components, out of which, cordycepin is considered most vital, due to its utmost therapeutic as well as nutraceutical potential. Moreover, the structure similarity of cordycepin with adenosine makes it an import antbioactive component, with difference of only hydroxyl group, lacking in the 30 position of its ribose moiety. Cordycepin is known for various nutraceutical and therapeutic potential, such as anti-diabetic, anti-hyperlipidemia, anti-fungal, anti-inflammatory, immunomodulatory, antioxidant, anti-aging, anticancer, antiviral, hepato-protective, hypo-sexuality, cardiovascular diseases,antimalarial,anti-osteoporotic,anti-arthritic,cosmeceutical etc. which makes it a most valuable medicinal mushroom for helping in maintaining good health. In this review, effort has been made to bring altogether the possible wide range of cordycepin’s nutraceutical potential along with its pharmacological actions and possible mechanism. Additionally,it also summarizes the details of cordycepin based nutraceuticals predominantly available in the market with expected global value. Moreover, this review will attract the attention of food scientists, nutritionists, pharmaceutical and food industries to improve the use of bioactive molecule cordycepin for nutraceutical purposes with commercialization to aid and promote healthy lifestyle, wellness and wellbeing.
... Antitumor, anti-diabetic, anti-inflammatory, antimicrobial, inhibit platelet aggregation, hypolipidemic, analgesic, immunomodulatory [15,[30][31][32] Adenosine ...
Article
Full-text available
Cordyceps is a rare naturally occurring entomopathogenic fungus usually found at high altitudes on the Himalayan plateau and a well-known medicinal mushroom in traditional Chinese medicine. Cordyceps contains various bioactive components, out of which, cordycepin is considered most vital, due to its utmost therapeutic as well as nutraceutical potential. Moreover, the structure similarity of cordycepin with adenosine makes it an important bioactive component, with difference of only hydroxyl group, lacking in the 3´ position of its ribose moiety. Cordycepin is known for various nutraceutical and therapeutic potential, such as anti-diabetic, anti-hyperlipidemia, anti-fungal, anti-inflammatory, immunomodulatory, antioxidant, anti-aging, anticancer, antiviral, hepato-protective, hypo-sexuality, cardiovascular diseases, antimalarial, anti-osteoporotic, anti-arthritic, cosmeceutical etc., which makes it a most valuable medicinal mushroom for helping in maintaining good health. In this review, effort has been made to bring altogether the possible wide range of cordycepin’s nutraceutical potential along with its pharmacological actions and possible mechanism. Additionally, it also summarizes the details of cordycepin based nutraceuticals predominantly available in the market with expected global value. Moreover, this review will attract the attention of food scientists, nutritionists, pharmaceutical and food industries to improve the use of bioactive molecule cordycepin for nutraceutical purposes with commercialization to aid and promote healthy lifestyle, wellness and wellbeing.
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Cordyceps is a precious herb in the world; it was discovered in the Tibetan Plateau of China. It has many good uses for human health as used to replenish the kidney and soothe the lung for the treatment of fatigue, night sweating, homosexualities, hyperglycemia, hyperlipidemia, asthenia after severe illness, respiratory disease, renal dysfunction and renal failure, arrhythmias, and other heart diseases, and liver disease. Due to the production of natural cordyceps exploited each year is increasingly scarce and depleted in recent times. Therefore currently, the cultivation of this herb has been applied to meet the higher demand of the market. Using microcontrollers to monitor and manage environmental parameters such as light, temperature and humidity, and soil moisture, we have produced medicinal substances from cordyceps of similar high quality as natural products.
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Cordyceps militaris (CM), a valuable edible and medicinal fungus, has been used as traditional medicine to treat health conditions, as well as hyposexuality in Asian societies for over a century. Due to the high demand, several artificial cultivation methods have been developed for their biological activities. In this study, CM was cultured on medium that contained white rice and silkworm pupae, and the levels of cordycepin and adenosine, as well as its aphrodisiac effects in diabetes-induced erectile dysfunction (DIED), were evaluated. Diabetic rats were induced by streptozotocin (STZ) injection and administered orally with CM (0.1, 0.5, and 1.0 g/kg BW/day) for 3 weeks. Diabetic rats in negative and positive control groups received vehicle and sildenafil citrate (5 mg/kg), respectively. Results showed the changes in mating behaviour in which mount latency and intromission latency were significantly increased in diabetic rats, compared with the normal control group. Diabetic rats also showed a significant reduction in intracavernosal pressure (ICP) response to cavernous nerve stimulation, sperm count, testosterone level, penile nitric oxide synthase (NOS), and testicular superoxide dismutase (SOD) activities, when compared to the normal control group. Administration of CM (0.1, 0.5, and 1.0 g/kg BW/day) reversed the effects of diabetes on the mating behaviour, and the ICP responses to electrical stimulation. Moreover, the levels of penile NOS, testicular SOD activities, testosterone, and sperm count were significantly increased, and testicular malondialdehyde (MDA) levels were significantly decreased in these treated diabetic rats. Diabetic rats treated with sildenafil showed a significant induction in intromission frequency and NOS and SOD activities, as well as a marked increase in ICP responses. These results suggest that CCM exerts its aphrodisiac effect, possibly through activating testosterone production and suppressing oxidative stress to enhance erectile function in diabetic rats. 1. Introduction Erectile dysfunction or ED has been signified as an inability of the male to achieve a penile erection, as part of the overall multifaceted process of male sexual function [1]. ED status can arise in adult men of all ages, as its prevalence and incidence are associated with aging. The prevalence of ED in young men has been estimated to be as high as 30%, in which diabetes mellitus (DM) either type 1 or type 2 has a well-established and strong association with ED [2, 3]. The pathophysiology of diabetes-induced erectile dysfunction (DIED) is multifactorial and several mechanisms of ED have been proposed in diabetic patients, including increased oxygen free radicals and impaired nitric oxide (NO) synthesis [4]. The chronic hyperglycemia can lead to endothelial dysfunction, which is manifested as the decreased bioavailability of NO, resulting in insufficient relaxation of vascular smooth muscle of the corpora cavernosal [5]. The current first-line therapy for diabetic ED is phosphodiesterase type 5 inhibitors (PDE5Is), such as sildenafil (Viagra®), tadalafil (Cialis®), and vardenafil (Levitra®). However, PDE5-Is has been shown to have some adverse effects, i.e., headache, abnormal vision, dyspepsia, flushing, nasal congestion, and back pain, which may impact negatively on patient’s lifestyle [6]. Alternative approaches, such as herbal medicine, have been adopted for sexual improvement for centuries. To date, many plants have been reported to possess aphrodisiac potential, and their effects on sexual behaviour have been validated [7]. In Asian countries, herbal supplements derived from Cordyceps species have been traditionally used as prosexual agent, one of which is Cordyceps militaris (CM), a valuable medicinal mushroom in the family Clavicipitaceae [8]. Nowadays, instead of harvesting from natural resources, CM has been cultivated using the artificial culture medium and similar bioactive contents and medicinal potential as wild Cordyceps has been reported [9]. Due to the growing demand, a number of culture techniques and the media formula to support growth of CM have been developed, and many bioactive ingredients have been isolated, such as adenosine, cordycepin, D-mannitol, polysaccharides, nucleosides, amino acid, essential oils, ergosterol peroxides, and xanthophylls [9–11]. Several scientific evidences related to the mechanisms and efficacy of this fungus, such as anticancer, antihypertensive, antioxidant, antiapoptotic, and hypoglycemic effects, have been reported [9, 10, 12–14]. Its positive effects in sexual function and testicular function have also been elucidated in young male rats [15], middle-aged rats [16], and aged male rats [17]. However, research on its bioactivity as prosexual agent in DIED is still scarce. The objective of the present study was to ascertain if CCM had aphrodisiac activity in STZ-induced diabetic rats and to elucidate the underlying mechanisms. 2. Materials and Methods 2.1. Preparation of CCM Cordyceps militaris was obtained from the Department of Agriculture (DOA) in Thailand. CCM was prepared at the Department of Agricultural Science, Faculty of Agriculture, Natural Resources and Environment, Naresuan University. In brief, the mycelia were cultivated with modified of potato dextrose agar (MPDA) medium under stable conditions at 22°C for 2 weeks. The resultant culture was transferred to potato dextrose broth (PDB) medium, which was then incubated on rotary shaker at 22°C for 2 weeks before transferring the mycelium to sterilized rice cultured medium that contained white rice and silkworm pupae. The fruiting bodies with the inoculums were kept in 12 : 12 h light-dark at 18°C, 60-70% humidity until the mycelium had transformed into the fruiting bodies primordia. Then, the flasks were maintained at 22°C, 80-90% humidity for 64 days before the fruiting bodies had been reaped and immediately frozen at -20°C until used. 2.2. Detection of Cordycepin and Adenosine in CCM with High-Performance Liquid Chromatography (HPLC) Fingerprint Analysis Cordycepin and adenosine in CCM were determined according to Huang et al. (2009) with some modifications. In brief, the fruiting bodies were dried in hot-air oven (55°C, 48 h). The dried samples were ground using a homogenizer, and 1.0 g of powder was added into 10 ml of methanol : water (50/50, ) and sonicated for 30 min, followed by centrifugation at 9,900 g for 15 min 2 times. The obtained supernatant was then filtered through a 0.45 μm filter membrane before injecting into the HPLC system (Shimadzu, Japan) with a column (Restek, Ultra IBD; , 5 μm particle size) set at 35°C. The mobile phase was a mixture of water and methanol (90 : 10; ) with the flow rate at 1 ml/min and a UV-vis detector at 254 nm. Five standard solutions of cordycepin and adenosine (Sigma Chemical, MO, USA) (20 μl) were prepared and injected into the HPLC to create standard calibration curves. 2.3. Animals Eight-week-old Sprague-Dawley rats used in this study were specific-pathogen-free (SPF) grade and were purchased from M-CLEA Bioresource Co., Ltd. (Samut Prakan, Thailand). Procedures involving animal subjects were approved by the Naresuan University Animal Care and Use Committee (NUACUC). All animals were handled in accordance with the Guidelines for the Care and Use of Laboratory Animals (National Research Council of Thailand) with an effort to minimize animal suffering. Rats were maintained under controlled temperature (22 ± 1°C) and relative humidity () with 12 : 12 hours of reverse light and dark cycle at Naresuan University Centre of Animal Research (NUCAR), which has been accredited by AAALACi. All rats were fed ad libitum a standard diet (CP No. 082; C.P. Company, Bangkok, Thailand) and allowed free access to reverse osmosis (RO) water. In order to induce diabetes, fifty male rats received a single intraperitoneal injection of STZ (60 mg/kg BW, i.p.) (Sigma-Aldrich, USA), which was dissolved in citrate acid buffer (pH 4.5). Ten male rats in the normal control group received only citrate buffer. After 72 h, fasting blood glucose (FBG) levels were checked using glucometer (Accu-Chek Performa, Roche). Rats with FBG levels higher than 200 mg/dl were used and divided into five groups: (I) DM control, (II) DM+CCM 0.1 g/kg, (III) DM+CCM 0.5 g/kg, (IV) DM+CCM 1.0 g/kg, and (V) DM+sildenafil 5 mg/kg. CCM were weighted and blended thoroughly with a blender and were given by daily oral gavage for 3 weeks. Sildenafil citrate was given only one time 30 min before mating behaviour test. 2.4. Surgical Procedure: Ovariectomy For prevention of pregnancy, female rats were subjected to bilateral oophorectomy surgery before beginning the sexual function assessment. Each adult female rat was anaesthetized by 1.5-2.0% isoflurane (Piramal Critical Cares, Inc., USA) combined with oxygen. The lower abdominal skin and muscle were opened vertically 1 cm, and the uterine horn was pulled out and ligated before removal of the ovary, one at a time. The uterine horn was returned to the peritoneal cavity, and the wound was closed in two layers (abdominal muscle and skin) using sterile sutures. The skin was then disinfected with povidone iodine and covered with Fixomull Stretch®. Each rat received an intramuscular tramadol (5 mg/kg) to ameliorate postoperative pain and allowed at least two weeks for full recovery. The ovariectomized female rats were artificially brought into oestrus phase by the administration of estradiol (0.025 mg/kg) and progesterone (1 mg/kg) at 48 and 4 hours before mating, respectively. 2.5. Mating Behaviour Assessment Mating tests were conducted in a custom made clear glass chamber . Each male rat was allowed to habituate in the chamber for 5 min before introducing a sexually receptive female rat into the chamber. The following male sexual behaviour parameters were calculated after monitoring for 30 min: (i)Mount latency (ML). The time interval between the introduction of the female and the first mount by the male(ii)Intromission latency (IL). The time interval between the introduction of the female and the intromission by the male(iii)Ejaculatory latency (EL). The time interval between the first intromission and ejaculation(iv)Mount frequency (MF). The number of mounts from the time of introduction of the female until ejaculation(v)Intromission frequency (IF). The number of intromissions from the time of introduction of the female until ejaculation(vi)Ejaculation frequency (EF). The number of ejaculations in a sexual cycle. Their behaviour was recorded with the digital VDO camera (LYD-808C, China) for offline analysis by two observers to ensure accuracy. 2.6. In Vivo Assessment of Erectile Function After completion of mating test, each male rat was anaesthetized with 2-2.5% isoflurane combination with oxygen. The ventilation rate, pulse rate, temperature, and heart rate were monitored via PhysioSuite® (Kent Scientific, USA). The carotid artery of rat was cannulated to measure mean arterial blood pressure (MAP) by using PowerLab® (AD Instruments, Australia). The penile skin was removed, and a polyethylene tube was inserted with heparinized saline via a 22-gauge needle for measuring intracavernosal pressure (ICP). The lower abdomen was opened exposing the cavernous nerve, which was then stimulated via a copper bipolar electrode connected to the PowerLab®. The cavernous nerve was stimulated by electrostimulation, starting from 0.25, 0.50, 0.75, 1, 2, 3, 4, and 5 to 10 volts at a frequency of 20 Hz for 60 sec for each voltage. The results were recorded by LabChart (version 7.3.7; AD Instruments, Australia) connected to a computer. Since electrostimulation slightly lowered MAP, ICP was normalized by MAP as ICP/MAP. Upon completion of experiment, rats were euthanized, and their penis and left testis were immediately collected and stored at -80°C for further analysis. The relative weights of penis and testis were calculated by the following formula: . 2.7. Determination of Serum Testosterone and Sperm Concentration and Motility Blood was collected from abdominal aorta and put into the nonanticoagulated tube and stored at 4°C before sending to the Biolab Medical Clinic, Phitsanulok, Thailand, for testosterone analysis. Semen was collected from the caudal epididymis and vas deferens and diluted in 1 M phosphate buffer saline (PBS) at 37°C. Then, 10 μl of sample was transferred into the Makler chamber and analysed under a light microscope. Sperm motility was recorded as video files for offline analysis. The number of sperm was counted and expressed as × 10⁶ per milliliter according to the WHO manual. 2.8. Measurement of Penile Nitric Oxide Synthase (NOS) Activity NOS activity in the penis was determined using the nitric oxide synthase assay kit (Cat. No 482702; Calbiochem®, Germany). The penile tissue was weighted and homogenized with ice-cold 1 M PBS (pH 7.4) by homogenizer (Ultra-turexT8, Germany). The sample was centrifuged at 10,000 g for 20 minutes and filtered through 0.45 μm membrane filter, and then the supernatant was obtained by centrifuging the sample at 100,000 g, 4°C for 15 min. The penile NOS activity was measured, following the manufacturer’s instructions. The absorbance at 540 nm was determined using a microplate reader (1401, LabSystem, Finland). Blank wells were used to normalize the yield. 2.9. Measurement of Superoxide Dismutase (SOD) Activity Testicular SOD was measured using the superoxide dismutase assay kit (Calbiochem®, cat #574601, Merck Millipore). Testis tissue was homogenized with buffer (10% ), pH 7.2 (containing 1 mM EGTA, 70 mM sucrose, 20 mM HEPES, and 210 mM mannitol). The homogenate was centrifuged at 1,500 g for 5 min. The supernatant was collected and centrifuged at 10,000 g, 4°C for 15 min. The SOD activity in the sample was then processed according to the manufacturer’s instructions and determined by a microplate reader with the absorbance at 450 nm. 2.10. Measurement of Thiobarbituric Acid Reactive Substances (TBARS) Concentrations of TBARS in testes tissues were measured according to the method of Ohkawa et al. [18] with some modifications. Testicular tissue was homogenized (10% ), in ice-cold 1 M phosphate buffer (pH 7.4). The homogenate was centrifuged at 4,000 g, 4°C for 15 min. Sample supernatant (100 μl) was added into the vial containing 1,500 μl of 20% acetic acid (pH 3.5), 200 μl of 8.1% sodium dodecyl sulphate (SDS), and 1,500 μl of 0.8% of thiobarbituric acid (TBA). The mixture was incubated for 60 min at 95°C and immediately cooled on ice, followed by centrifugation at 10,000 g for 3 min. The resulting supernatant was then used as the enzyme source for the determination of the malondialdehyde (MDA) level by the optical density (OD) measurement of the pink complex at 532 nm. MDA values were calculated using tetramethylpiperidine (TMP) as a standard curve and expressed as nmol/mg of protein that determined absorbance at a wavelength of 562 nm. Protein level was measured using the Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific, USA). 2.11. Histological Examination of Testicular Tissues The right testis was fixed in 10% neutral buffered formalin over 24 hours. The tissues were then processed with 70-100% ethanol and xylene, respectively. The infiltrated tissues were embedded in paraffin blocks and cut at 2 μm by using semiautomatic microtome (Leica, Germany). All slides were stained with hematoxylin and eosin (H&E), and histological changes were observed by light microscopy. 2.12. Statistical Analysis The value of mating parameters, sperm concentration, NOS and SOD activities, and MDA level were presented as . ICP, MAP, and testosterone levels were presented as . Data were analysed using one-way analysis of variance (ANOVA) followed by Dunn’s post-hoc Multiple Comparison Test (Graph Pad Prism 7.4, GraphPad Software, San Diego, USA). values of less than 0.05 were regarded as statistically significant. 3. Results and Discussion 3.1. Cordycepin and Adenosine Contents in CCM Many species of Cordyceps are being cultivated in artificial medium for their medicinal and pharmaceutical properties. Cordycepin, a derivative of the nucleoside adenosine, has been shown to be the first and is the main active constituent isolated from Cordyceps sp. [9]. In this study, we performed an analysis of bioactive compounds and found that the adenosine and cordycepin contents in CCM were and , respectively (Figure 1). This amount of cordycepin is relatively similar to that cultured in silk worm pupae medium () and higher than that obtained in brown rice medium (), as reported by Kang et al. [19]. However, cordycepin production could yield with a maximum of about 445 mg/l, when CM was cultured in submerged conditions [20].
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