THERAPEUTIC PROPERTIES AND CURRENT MEDICAL USAGE OF MEDICINAL MUSHROOM: GANODERMA LUCIDUM

THERAPEUTIC PROPERTIES AND CURRENT MEDICAL USAGE OF MEDICINAL MUSHROOM: GANODERMA LUCIDUM

1. Deepalakshmi and S. Mirunalini*

Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalai Nagar, Tamil Nadu, India

ABSTRACT

Mushrooms are an important natural source of food and medicines. Traditionally medicinal properties of mushroom have been well demonstrated particularly in Eastern Asian countries. In modern clinical practices, the bioactive compounds derived from the extract of mushroom sporocarps or mycelium has been widely used for the prevention and treatment of various human diseases such as cancer, diabetes, immune system disorders and infections. Recently, considerable attention is focused on anticarcinogenic bioactive compounds particularly those derived from medicinal or wild mushrooms. The present review analyses the potential therapeutic properties of medicinal mushroom Ganoderma lucidum and their applications in human health care.

β-D- glucans

INTRODUCTION: In recent years, natural products have attracted extensive attention in drug discovery and development ¹. However, there is strong and consistent evidence showing that a diet rich in natural product such as fruits, vegetables, herbs, cereals, sprouts and edible mushrooms are associated with decreased risk of many diseases ². Such natural bioactive substances possess an enormous structural and chemical diversity, unsurpassable by any synthetic library; they are evolutionally optimized as drug-like molecules and might be considered biologically validated.

It is empirically known that mushrooms have been valued throughout the world as both food and medicine for thousands of years. They represent a major and as yet largely untapped source of potent pharmaceutical products. There is a common saying that “medicines and food have a common origin” ³. However, dietary mushrooms provide a wide variety of medicinal properties and they are effective against certain life-threatening diseases. It is estimated that there are approximately 1.5 million species of fungi in world, of which approximately 82,000 are described ⁴. About of the known species belong to macrofungi, of which about 5,000 are edible and 2,000 safe ⁵. Fungi from the basidiomycota division are of great interest due to the presence of a large number of biological active compounds they contain ⁶.

Traditionaly, Ganoderma is highly regarded as a herbal treatment and is claimed to alleviate or cure virtually all diseases ^{5, 7, 8}. Ling Zhi encompassed several Ganoderma species,  which are widely used for medicinal purposes eg., Ganoderma lucidum, Ganoderma luteum steyaert, Ganoderma atrum Zhao, Xu and Zhang, Ganoderma applanatu (pers.:Wallr) pat.,Ganoderma australe (Fr.) pat., Ganoderma capense (Lloyd) Teng, Ganoderma tropicum (Jungh) Bres.,Ganoderma tenue Zhao, Xu and Zhang and Ganoderma sinense Zhao, Xu and Zhang. Worldwide, more than 250 Ganoderma species have been described ^{9, 10}. However, in therapeutic practices and literature citation, Ganoderma usually refers to the species of Ganoderma lucidum.

Ganoderma lucidum, is a Woody basidiomycotina mushroom belonging to the family of Ganodermaceae of polyporales, which is widely used in oriental medicine for longevity and health promotion ¹¹. Ganoderma lucidum is commonly named as “Lingzhi’’ in china, “Youngzhi’’ in Korea, “Reishi’’ in Japan, and just “Ganoderma’’ in USA ¹². Ganoderma lucidum is the annular mushroom grows in a wide variety of dead or dying trees, eg., Deciduous trees especially oak, maple, elm, willow, sweet gum, magnolia and locust and less frequently found on coniferous tree (eg., Larix, ptea, pinus) in Europe, Asia and North and South America (in temperate rather than subtropical region ¹³.

Current research is focused on purification and characterization of the bioactive components and determination of their clinical values ^{5, 7, 8}. Chemical investigations on the fruiting bodies, spores and mycelia of Ganoderma lucidum reveal that they contain various bioactive substances ^14. Because of its presumed health benefits and apparent absence of side effects; it has attained a reputation in the east as the ultimate herbal substance. Ling Zhi has now been added to the American Herbal Pharmacopoeia and Therapeutic compendium.

Bioactive Constituents: The bioactive constituent of Ganoderma lucidum originates from its chemical composition. The mushroom contains polysaccharides, polysaccharide- peptide complex, β-glucans, lectins, organic germanium(Ge), adenosine, triterpenoids, phenols, steroids, amino acids, lignin, mycins, vitamins, nucleotides and nucleosides each having their own outstanding medicinal effects ^{15, 16}. Most fractions of identified polysaccharides and triterpenes have more than hundred compounds that are potent immune-modulators, antioxidants and /or chemo preventive and tumoricidal ¹⁷.

Nutritional Values: Nutritional analysis of several mushroom species of different origins had been carried out in many laboratories in the world. But nutritional values of locally cultivated mushrooms remain speculative. Moreover, nutritional composition is affected by many factors; these include differences among strains, composition of growth substrate, method of cultivation, stage of harvesting, specific portion of the fruiting bodies used for analysis ¹⁸. The composition of Ganoderma lucidum extract (% of dry weight) Table 1, consisted of folin-positive material (68.9%), glucose (11.1%), protein (7.3%) and metals (10.2%) (K, Mg and Ca are the major components with Ge having the 5^th highest metal concentration at (489µg/g). These results generally agree with those reports of other authors ^{13, 12, 22}. Whereas, nutritional analysis of Ganoderma lucidum contains mainly protein, fat, carbohydrate Table 2 and fiber.

TABLE 1: MAIN NUTRITIONAL COMPONENT OF FERMENTED PRODUCTS OF GANODERMA LUCIDUM AND A NON-FERMENTED CONTROL ON A DRY BASIS ²³

The starch content of the fermented control reached 64.5%, the reducing sugar content was only 4.2%. However, the starch content of the fermented product decreased significantly (p<0.001) from 64.5 to 25.3%; while the reducing sugar content increased significantly (p<0.001) from 4.2 to 20.6%. Solid state fermentation (SSF) also produced a significantly (p<0.01) increased from 11.0 to 16.5% in protein content. After SSF, the crude fat content decreased significantly (p<0.01). However, these are qualitative and quantitative differences in the chemical composition of Ganoderma lucidum product depending on the strain, origin, extracting process and cultivation conditions.

TABLE 2: CARBOHYDRATE COMPOSITIONS OF CRUDE GANODERMA LUCIDUM EXTRACT ¹⁹

Other Constituents: Reishi also contain sterols, amino acids Table 3 Soluble  proteins, oleic acid, cyclo-octa sulfur, an ergosterol peroxide (5,8-epidioxy-ergosta-6,22E-dien-3-ol) and the cerebrosides(4E’,8E)-N-D-2’-hydroxystearoyl-1-0-β-D-glucapyran-ocyl-9-methyl-4-8-sphingadienine and (4E,8E)-N-D-2’-hydroxypamitoyl-1-D-β-D-glucopyranosyl-9-methyl-4-8-sphingadienine ^{9, 10, 17, 18,20}. Regarding the inorganic ions, the mushroom contains Mg, Ca, Zn, Mn, Fe, Cu and Ge. The spores themselves contain choline, ketain, tetra cosanoic acid, palmitic acid, ergosta-7, 22-dien-3-ol, nonadecanoicacid, behenicacid, tetracosane, hentriacontane, ergosterol and β-sitosterol. One of the lipids isolated from Ganoderma lucidum is pyrophosphosphatidic acid ^{10, 15, 17, 24}.

TABLE 3: AMINO ACID ANALYSES OF GANODERMA LUCIDUM EXTRACT ¹⁹

Therapeutic Applications:

Preclinical and Clinical Studies:

Ganoderma lucidum has also become popular because of its promising properties that might extend life span while increasing vigor and vitality ⁵⁸. Ganoderma lucidum has been reported to have a number of pharmacological effect including immunomodulating, antiartherosclerotic, anti-inflammation, analgesic, chemopreventive, anti-tumor, radioprotective, sleep-promoting, antibacterial, antiviral (including anti-HIV), hypolipidemic, anti-fibrotic, hepatoprotective, diabetic, antioxidative and radical-scavenging, anti-aging, hypo-glycemic and anti-ulcer properties Table 4 ^{10, 22, 24, 26, 27}.

TABLE 4: THERAPEUTIC EFFECTS AND BIOACTIVE COMPOUNDS OF GANODERMA LUCIDUM

Ganoderma lucidum has now become recognized as an alternative adjuvant in the treatment of leukemia, carcinoma, hepatitis and diabetes ^{6-8, 17}. Since the last decades, clinical trials on the use of Ganoderma lucidum preparation used to treat cancer and other diseases have been reported in international peer-reviewed journals.

Immunomodulatory Activtiy: Immunomodulatory properties alone with low cytotoxicity raise the possibility that it could be effective in the cancer patients receiving conventional chemotherapy and/or radiational treatment, to build up immune resistance and decreased toxicity. Numerous experimental and clinical investigations demonstrated that Ganoderma lucidum had immunomodulatory activities. A number of reports have demonstrated that Ganoderma lucidum polysaccharides stimulated immune function both in vivo and in vitro. Recent literature has been found that Ganoderma lucidum modulate many components of the immune system such as the antigen-presenting cells, NK cells, T and B lymphocytes, macrophages, resulting  in the production of cytokines, including interleukins, tumor necrosis factor-α (TNF-α) and interferon ²⁸. Chen W C et al., (1995) showed that a crude aqueous extract of Ganoderma lucidum probably administered was effective in enhancing the recovery of leukocytes count, splenic blastogenic responses and splenic CD4 and CD8 T cell subsets in mice subjected to γ-irradiation ²⁹. Choi-Lan-Ha et al., 2003 have demonstrated the inhibitory effect of the Chinese herb Ganoderma lucidum mycelium on gut immunoglobulin A responses to cholera toxin in mice ³⁰.

Mechanisms Of Ganoderma Lucidum On Immunomodulatory Activity: The immunomodulating effect of Ganoderma lucidum were extensive, including promoting the function of antigen-presenting cells, mononuclear phagocytes system, humoral immunity and cellular immunity and cellular immunity and the action site of Ganoderma lucidum was speculated to be located in the course of proliferation and differentiation of immune precursor cells to effector cells.

Mono-Nuclear Phagocyte System: Many scientific investigations have found that one polysaccharide isolated from Ganoderma lucidum which were mainly composed of β-D-glucans Fig. 1 could TNF-α Synthesis in primary culture of human peripheral blood mononuclear cells (PBMC). And exposure of human neutrophils to Ganoderma lucidum  polysaccharides time-dependently caused increases in protein kinase C (PKC), P³⁸ mitogen-activated protein kinase (MAPK), hematopoietic cell kinase (HCK), moreover tyrosine kinase Lyn activities, these may be the action that corresponded to an enhanced unspecific immune function ³¹.

FIG. 1: POSSIBLE REPEATING UNIT OF GANODERMA LUCIDUM GLUCANS ³²

Antigen-Presenting Cells: Dendritic cells (DC), a kind of professional antigen- presenting cells, are pivotal for initiation of primary immune response. Cao LZ et al., reported that Ganoderma lucidum  promote not only the maturation of cultivated murine bone marrow derived DC in vitro but also the immune response initiation induced by DC ³³.

Natural Killer (Nk) Cells: Recent report suggested that treatment with the water soluble extract of Ganoderma lucidum (F3) F3-Polysaccharide fraction could increase the presence of natural killer cells (CD56 (+) marker) significantly from 1.1% to 3.2% in mononuclear cells, indicating that F3 quantitatively influenced NK cells activities ³⁴.

T-Lymphocytes: The cell–mediated immune function was also enhanced by Ganoderma lucidum, as suggested by the observation that Ganoderma lucidum promoted the mixed lymphocyte reaction (MLC) ³⁵. It also exerted an increasing effect on the induction of delayed hypersensitivity to protein antigen. Ganoderma lucidum polysaccharides like BN3A, BN3B and BN3C are three kinds of polysaccharides significantly increased the lymphocyte proliferation induced by Con A and IL-2 production in the normal mice, as well as in the aged mice in vitro. BN3A and BN3C also could antagonize the suppressive effect of hydrocortisone on the proliferation of mouse spleen cells ³⁶. Moreover, Ganoderma lucidum increased the production of IFN-γ and significantly increase IFN-γ mRNA expression in the T-Lymphocytes ³⁷.

B-Lymphocytes: A bioactive fraction (GLIS), isolated from the fruiting body of Ganoderma lucidum could stimulate the activation, proliferation and differentiation of B-Lymphocytes. The B-Lymphocytes were to enlarged, expressed CD71 and CD25 on the cell surface, and showed an increase in the secretion of immunoglobulin. However, the activation of B-Lymphocytes by GLIS did not depend on the activation of T-Lymphocytes. It was associated with stimulation of the expression of protein kinase C α and protein kinase C γ in B-Lymphocytes by GLIS directly ³⁸.

Macrophages: Macrophages are responsible for killing pathogen in the body. The substances from Ganoderma lucidum activation of macrophages which result in the release of cytokines and other mediators. Whereas, polysaccharides from Ganoderma lucidum in particular β-D-glucans are potent stimulators of murine and human macrophages in vitro and in vivo ³⁹. Crude water- extracted polysaccharides isolated from fresh fruiting bodies of Ganoderma lucidum potentiated the production of cytokines, including IL-1β, IL-6, IFN-γ and TNF-α by human macrophages, which were anti-proliferative, differentiation and apoptosis inductive to the HL-60 and the U937 leukemic cells.

Anti-inflammatory Effects: Anti-inflammatory drug make up about half of analgesics, remedying pain by reducing inflammation as opposed to opioids which affect the brain. Joseph et al., showed the anti-inflammatory activity of chloroform extract of Ganoderma lucidum in dose dependent carrageenan induced acute and format induced chronic inflammatory models in mice ⁴⁰.  In addition Akihisa et al., also investigated the anti-inflammatory activity of GA-A, -F, -DM   and   -T-Q in 1-O-tetradecanoylphorbol 13 –acetate- induced inflammation in mice ⁴¹.

Anticancer Activities: Ganoderma lucidum, an oriental medical mushroom has been used widely in Asian countries for centuries to prevent or treat different diseases including cancer. Dried powder of Ganoderma lucidum, which was recommended as a cancer chemotherapy agent, is currently used popularly worldwide in the form of dietary supplements ⁴².

Ganoderma lucidum extracts were reported to possess cytotoxic activity against various cancer cell lines including leukemia, lymphoma, multiple myeloma ^{43, 44} and human breast cancer MCF-7 ⁴⁵. The cytotoxic effect of   Ganoderma lucidum as demonstrated by the studies of Jiang et al., and Zhu et al., in a concentration dependent manner ^{46, 47}. This activity of Ganoderma lucidum can be attributed directly to specific compounds from experiments employing isolated and purified molecules.

Wang S. Y. et al., showed that the anti-tumor effect of Ganoderma lucidum was mediated by cytokines released from activated T-Lymphocytes and macrophages ³⁹. Ganoderma lucidum could potentiate the production of cytokine including interleukin-1, interleukin-6, tumor necrosis factor, and interferon in which two antitumor cytokines, tumor necrosis factor and interferon, acted synergistically on the inhibition of leukemic-cell growth and markedly induced leukemic - cell apoptosis. The organic Germanium in Ganoderma lucidum may also contribute to its anti-tumor activity ⁴⁹. W. Tang et al., (2006) proposed that GA-T may be a natural potential apoptosis-inducing agent for highly metastatic lung tumor and it may be also applied to treat other tumor cell lines Fig. 2.

FIG. 2: PROPOSED PATHWAY INVOLVED IN APOPTOSIS INDUCED BY GA-T ⁵⁰

Antioxidant Activities: An anti-oxidant may be defined as “any substance that when present at low concentrations, compared with those of the oxidizable substrate significantly delays or inhibits oxidation of that substrate” ⁵¹. Antioxidative effect of sporocarp extracts have not been examined so far. In as much as antioxidative activities have significant therapeutic effects; the fungal species could be used in therapy for a variety of disease states and in health nutrition as a source of naturally derived antioxidants.

These are easily noticed, collected and recognized in field and their secondary metabolites can be easily identified and extracted ⁵². Many literatures have demonstrated that chloroform extract of Ganoderma lucidum possesses significant capacity to inhibit free radical formation and scavenging activity. The chloroform extract showed significant superoxide scavenging activity (IC₅₀: 144.6±1.5 µg / ml) and is of potential interest as source of strong natural antioxidant in the food and cosmetics industries. GA-A, -B, -C and -D also showed antioxidative effect against pyrogallol-induced erythrocyte membrane oxidation and Fe (II) ascorbic acid –induced lipid peroxidation ⁵³.

Anti-aging Effect: Relatively recent research on anti-ageing effect from natural products are of the highest importance for medical stakes (Oncology and immunology) and industrial development (Pharmacy and Cosmetics), it also opens prospects to scientific screening of antioxidizing natural agents among higher fungi ⁵⁴.

The oxidative damage caused by these free radicals may be related to ageing and diseases, such as atherosclerosis, diabetes, cancer and cirrhosis ⁵⁵. However, antioxidant supplements or food containing antioxidants may be used to reduce oxidative damage, by not only providing essential vitamins and minerals, but include important chemo-protective agent capable of protecting against some forms of cancer⁵⁷. Guesnet et al, 2003 stated that GA-B,-C2 and –G have anti-aging effect and can be used as cosmetic agents in various forms ⁵⁶.

Antidiabetic Effect: The polysaccharide factions of Ganoderma lucidum have potential hypoglycemic and hypolipidemic activities have been demonstrated by animal studies. However to evaluate the anti-diabetic efficacy and safety, a clinical study were carried out on polysaccharide fractions extraction from Ganoderma lucidum (Ganopoly) by a patented technique in 71 patients with confirmed type II diabetes mellitus(DM) was carried out ⁶⁰. This study demonstrated that Ganopoly was well tolerated, efficacious and safe in lowering blood glucose concentrations.

Antibacterial Effects: Several studies have been demonstrated that Ganoderma lucidum contained antibacterial constituents that are able to inhibit gram-positive and gram-negative bacterias ^{61, 62}. The Aqueous extract of the carpophores of Ganoderma lucidum inhibited 15 types of gram-positive and gram-negative bacteria. However, further studies indicated that combination of Ganoderma lucidum extract with four antibiotics like ampicillin, cefazolin, oxytetracyline and chloramphenicol resulted in additive effects. Whereas, synergism in two instances where combined with cefazolin against Bacillus subtilis and Klebsiella oxytoca and antagonism in two instances ⁶³.

Commercialization: Ganoderma lucidum is usually prescribed in various forms. It may be injected as a solution of powdered spore. Under the name of LingZhi or Reishi, a number of Ganoderma lucidum  products are sold as over the counter products in the forms of health drinks, soup, syrup, tea, tablets, capsules, tincture or bolus (powdered medicine in honey) ^{61, 60}. In 2008, the worldwide production of Ganoderma lucidum was approximately 9500 tones, of which china contributed 6000 tones. Ganoderma lucidum is mainly used as a tonic and a remedy for the treatment of a variety of diseases. Hence, it may be noted that Ganoderma lucidum is an excellent source of medicine which is enabling the present need of human mankind in every aspect ⁵⁸.

CONCLUSION: As seen through the review, Ganoderma lucidum present several mechanism of action to develop its large number of therapeutical function. Over all evidence show that Ganoderma lucidum a mushroom of biomedical importance, contains a number of bioactive components, many of the biological response modifiers which activate our immune system for a multitude of defensive functions. However, immune-modulating effects of Ganoderma lucidum are associated with its anti-tumor activity. Likewise, the multiple mechanisms in chemoprevention activity of Ganoderma lucidum, as superoxide scavenging, lipid peroxidation is inhibiting and nitric oxide scavenging and anti-oxidation may be one of the important mechanism by which Ganoderma lucidum exerts its tumor inhibitory effect.

Even though, Ganoderma lucidum is an immortal golden medicinal fungus are still to be exploited commercially. The reason that some of the Ganoderma lucidum preparations are not available as medicines may be due to difficulties related to mass production. And, the majority of research programs had been focused on extracts from the fruiting body and there have been fewer studies on extracts from the cultivated fungi. Therefore, further research may be oriented in that direction. Since, most of the therapeutic effects of Ganoderma lucidum are based on in vivo and in vitro studies, clinical trials are needed to fully realize its potential.

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