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PubMed2001

Structural and immunological studies of a major polysaccharide from spores of Ganoderma lucidum (Fr.) Karst.

A polysaccharide isolated from spores of the fungus, Ganoderma lucidum, was found to be a complex glucan. On the basis of compositional and methylation analyses, periodate oxidation, Smith degradation, 1D and 2D NMR, and ESIMS experiments of the native polysaccharide and its degraded products, the polysaccharide was shown to have a backbone of beta-(1-->3)-linked D-glucopyranosyl residues, with branches of mono-, di- and oligosaccharide side chains substituting at the C-6 of the glucosyl residues in the main chain. Conformational analysis in aqueous solution and immunological activities of the native and degraded glucans were also investigated. The results suggested that the degree of substitution on the main chain and the length of side chains may be very important factors in determining the conformation and the biological activities of beta-(1-->3)-linked glucans.

Animals↗

PubMed2001

Chemical modifications of the (1-->3)-alpha-D-glucan from spores of Ganoderma lucidum and investigation of their physicochemical properties and immunological activity.

A linear (1-->3)-alpha-D-glucan was isolated from the spores of Ganoderma lucidum (Fr.) Karst. Six different functionalized derivatives of the (1-->3)-alpha-D-glucan-aminopropylated, hydroxyethylated, sulfated, carboxymethylated, carboxymethylated and sulfated, and benzylamidated-carboxymethylated-with varying degrees of substitution were synthesized. The structural features and physicochemical properties of all derivatives were investigated by means of chemical and spectral analyses, and their effects on lymphocyte proliferation and antibody production were tested in vitro and in vivo. In general, the structural and physicochemical properties, and lymphocyte proliferation activity of all samples varied with the functionalized groups and the degree of substitution. The results of immunological assays indicated that some modified derivatives had potent stimulating effects on lymphocyte proliferation and antibody production and the introduction of carboxymethyl group with low degree of substitution (DS<0.28) was the best choice on the improvement of the immunostimulating activity.

Adjuvants, Immunologic↗

PubMed2002

Activation of B lymphocytes by GLIS, a bioactive proteoglycan from Ganoderma lucidum.

A bioactive fraction (GLIS) was isolated from the fruiting body of the fungus Ganoderma lucidum using successive chromatographic steps. GLIS is a proteoglycan and has a carbohydrate: protein ratio of 11.5 : 1. The carbohydrate portion is composed of seven different monosaccharides, predominantly D-glucose, D-galactose and D-mannose in the molar ratio of 3.0 : 1 : 1.GLIS stimulated the proliferation of mouse spleen lymphocytes, resulting in a three to four-fold increase in the percentage of B cells. GLIS also activated mouse spleen lymphocytes, and most of the activated cells were B cells. The B cells were enlarged, expressed CD71 and CD25 on the cell surface, and showed an increase in the secretion of immunoglobulin. Lymphocytes also showed a slightly increased production of IL-2, whereas the secretion of IL-4 was not influenced by GLIS. Furthermore, GLIS did not influence the intracellular Ca2+ concentration of lymphocytes, but it enhanced the expression of protein kinase C alpha and protein kinase C gamma in B cells. According to our results GLIS is a new B cell-stimulating factor.

Animals↗

PubMed2002

Mechanism of the antiulcerogenic effect of Ganoderma lucidum polysaccharides on indomethacin-induced lesions in the rat.

Many cytokines, in particular tumor necrosis factor (TNF)-alpha have been known to play an important role in the pathogenesis of gastric mucosal lesions caused by various factors such as drugs and Helicobacter pylori infection. Our previous studies have shown that the polysaccharide fractions isolated from the fruiting bodies of Ganoderma lucidum (GLPS) prevented indomethacin- and acetic acid-induced gastric mucosal lesions in the rat. However, the mechanisms remain unclear. This study aimed to investigate whether GLPS had a direct mucosal healing effect in the indomethacin-treated rat, and to explore the possible mechanisms by determining the gastric mucosal mRNA and protein levels of TNF-alpha and ornithine decarboxylase (ODC) activity. In addition, the effects of GLPS on the cellular proliferation, ODC and c-Myc protein expression and mucus synthesis in the rat gastric cell culture (RGM-1) were examined. The present study demonstrated that GLPS at 250 and 500 mg/kg by intragastric input caused ulcer-healing effect in the rat; this was accompanied with a significant suppression of TNF-alpha gene expression, but with an increased ODC activity. In RGM-1 cells, GLPS at 0.05, 0.25 and 1.0 mg/ml significantly enhanced [3H]thymidine incorporation and ODC activity in a concentration-dependent manner. However, these effects were abrogated by the addition of the ODC inhibitor, DL-alpha-difluoromethyl-ornithine (DFMO). GLPS at 0.25-1.0 mg/ml also increased mucus synthesis, as indicated by the increased D-[6-3H]glucosamine incorporation in RGM-1 cells. Furthermore, GLPS at 0.05-1.0 mg/ml increased the c-Myc protein expression. These findings indicated that GLPS produced a mucosal healing effect in the rat model, perhaps due partly to the suppression of TNF-alpha and induction of c-myc and ODC gene.

Animals↗

PubMed2003

Triterpene-enriched extracts from Ganoderma lucidum inhibit growth of hepatoma cells via suppressing protein kinase C, activating mitogen-activated protein kinases and G2-phase cell cycle arrest.

The medicinal mushroom Ganoderma lucidum (G. lucidum) has been used in the Orient for the prevention and treatment of various diseases including cancer. Except for the immune enhancing properties of its polysaccharide constituent, very little is known about the anticancer activity of another major constituent, triterpenes. In this report, we studied the anticancer mechanism of triterpene-enriched extracts from G. lucidum. The triterpene-enriched fraction, WEES-G6, was prepared from mycelia of G. lucidum by sequential hot water extraction, removal of ethanol-insoluble polysaccharides and then gel-filtration chromatography. We found that WEES-G6 inhibited growth of human hepatoma Huh-7 cells, but not Chang liver cells, a normal human liver cell line. Treatment with WEES-G6 caused a rapid decrease in the activity of cell growth regulative protein, PKC, and the activation of JNK and p38 MAP kinases. The changes in these molecules resulted in a prolonged G2 cell cycle phase and strong growth inhibition. None of these effects were seen in the normal liver cells. Our findings suggest that the triterpenes contained in G. lucidum are potential anticancer agents.

Carcinoma, Hepatocellular↗

PubMed2002

Structural features of immunologically active polysaccharides from Ganoderma lucidum.

Three polysaccharides, two heteroglycans (PL-1 and PL-4) and one glucan (PL-3), were solubilized from the fruit bodies of Ganoderma lucidum and isolated by anion-exchange and gel-filtration chromatography. Their structural features were elucidated by glycosyl residue and glycosyl linkage composition analyses, partial acid hydrolysis, acetolysis, periodate oxidation, 1D and 2D NMR spectroscopy, and ESI-MS experiments. The data obtained indicated that PL-1 had a backbone consisting of 1,4-linked alpha-D-glucopyranosyl residues and 1,6-linked beta-D-galactopyranosyl residues with branches at O-6 of glucose residues and O-2 of galactose residues, composed of terminal glucose, 1,6-linked glucosyl residues and terminal rhamnose. PL-3 was a highly branched glucan composed of 1,3-linked beta-D-glucopyranosyl residues substituted at O-6 with 1,6-linked glucosyl residues. PL-4 was comprised of 1,3-, 1,4-, 1,6-linked beta-D-glucopyranosyl residues and 1,6-linked beta-D-mannopyranosyl residues. These polysaccharides enhanced the proliferation of T- and B-lymphocytes in vitro to varying contents and PL-1 exhibited an immune-stimulating activity in mice.

Adjuvants, Immunologic↗

PubMed1997

A lectin from mycelia of the fungus Ganoderma lucidum.

A lectin (GLL-M) was isolated from mycelia of Ganoderma lucidum using affinity chromatography on BSM-Toyopearl. GLL-M is a monomer in its native form with a M(r) of 18,000. Another lectin was also purified from fruiting bodies of the same fungus. The two lectins were partially compared with each other.

Amino Acids↗

PubMed1998

Anti-HIV-1 and anti-HIV-1-protease substances from Ganoderma lucidum.

A new highly oxygenated triterpene named ganoderic acid alpha has been isolated from a methanol extract of the fruiting bodies of Ganoderma lucidum together with twelve known compounds. The structures of the isolated compounds were determined by spectroscopic means including 2D-NMR. Ganoderiol F and ganodermanontriol were found to be active as anti-HIV-1 agents with an inhibitory concentration of 7.8 micrograms ml-1 for both, and ganoderic acid B, ganoderiol B, ganoderic acid C1, 3 beta-5 alpha-dihydroxy-6 beta-methoxyergosta-7,22-diene, ganoderic acid alpha, ganoderic acid H and ganoderiol A were moderately active inhibitors against HIV-1 PR with a 50% inhibitory concentration of 0.17-0.23 mM.

Anti-HIV Agents↗

PubMed2000

Effect of fatty acids on the mycelial growth and polysaccharide formation by Ganoderma lucidum in shake flask cultures.

Fatty acids were added into the media to investigate their effects on the mycelial growth and polysaccharide formation by Ganoderma lucidum. The experiments were carried out in freely suspended cultures or immobilized cultures using shake flasks. The results indicate that the extent of stimulation or inhibition were associated with the types and levels of fatty acids. Oleic acid at the level of 0.15 g/100 ml led to a significant increase in cell concentration from 0.20 to 0.46 g/100 ml in a suspended culture and palmitic acid was of great advantage to polysaccharide production. In contrast, linoleic acid (0.1 g/100 ml) drastically suppressed both mycelial growth and polysaccharide formation. In immobilized cultures with fatty acids, the stimulation of mycelial growth remained the same level, but the enhancement of polysaccharide production became less. In addition, the growth of G. lucidum in the pattern of immobilization might be beneficial to the production of mycelia and polysaccharide.

Journal Article↗

PubMed2003

Characterization of a polysaccharide-protein complex from Ganoderma tsugae mycelium by size-exclusion chromatography combined with laser light scattering.

A water-soluble polysaccharide-protein complex (GM3) extracted from the mycelium of Ganoderma tsugae was characterized using size-exclusion chromatography combined with laser light scattering (SEC-LLS). Two peaks coded as fractions I and II appeared in the SEC pattern of GM3 in 0.5 M NaCl aqueous solution, corresponding to the weight-average molecular mass (M(w)) of 355 x 10(4) and 6.3 x 10(4), respectively. The relationship between the radius of gyration ( (z)(1/2)) and M(w) showed that molecules of fraction I exhibited more compact coil conformation than that of fraction II in 0.5 M NaCl aqueous solution at 25 degrees C. To clarify the component of polysaccharide and protein in each fraction, the sample GM3 was treated with 0.2 M NaOH aqueous solution to degrade polysaccharide and trypsin to hydrolyze protein. The obtained products were analyzed by SEC combined with detectors such as UV, differential refractive index (DRI) and LLS. The results indicated that both the fractions I and II were protein-bound polysaccharide, but had different protein content and degree of branching, resulting in the difference of the chain conformation.

Basidiomycota↗

PubMed2002

Regulation on maturation and function of dendritic cells by Ganoderma lucidum polysaccharides.

Ganoderma lucidum polysaccharides (Gl-PS) exhibits a variety of immunomodulatory activities, and dendritic cells (DC) are professional antigen presenting cells, which are pivotal for initiation of primary immune response. In this study, the regulatory effects of Gl-PS on maturation and function of cultured murine bone marrow derived DC were investigated in vitro. Gl-PS (0.8, 3.2, or 12.8 microg/ml) could increase the co-expression of CD11c and I-A/I-E molecules on DC surface, promote mRNA expression of cytokine IL-12 p40 in DC and augment protein production of IL-12 P40 in culture supernatants. The lymphocyte proliferation of mixed lymphocyte culture (MLC) induced by mature DC was enhanced by Gl-PS, either. Gl-PS was shown to promote not only the maturation of cultured murine bone marrow derived DC in vitro, but also the immune response initiation induced by DC.

Animals↗

PubMed2002

Antitumor activity of the sporoderm-broken germinating spores of Ganoderma lucidum.

The inhibitory effects of the dormant spores, the germinating spores, the sporoderm-broken germinating spores (SBGS), and the lipids extracted from the germinating spores of Ganoderma lucidum on the growth of mouse hepatoma, sarcoma S-180, and reticulocyte sarcoma L-II cells were investigated, respectively. The dormant spores could be activated by germination, and thus the bioactivities of the spores might be enhanced. The sporoderm-broken spores could show much higher bioactivities than the whole spores. Both the lipids extracted from the germinating spores and the SBGS of G. lucidum had remarkable antitumor effects in a dose-dependent manner, and could significantly inhibit three tumors with an inhibition of 80-90%.

Administration, Oral↗

PubMed2000

Antiherpetic activities of acidic protein bound polysacchride isolated from Ganoderma lucidum alone and in combinations with acyclovir and vidarabine.

To investigate antiherpetic activity, an acidic protein bound polysaccharide (APBP) was isolated from carpophores of Ganoderma lucidum. This brownish APBP was isolated from water soluble substances of the carpophores by activity-guided isolation method. APBP was tested for its antiviral activity against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) by plaque reduction assay in tissue culture. APBP showed potent antiviral activity against HSV-1 and HSV-2 in Vero cells at its 50% effective concentration (EC(50)) of 300 and 440 microg/ml, respectively. APBP had no cytotoxicity on Vero cells at a concentration of 1 x 10(4) microg/ml. APBP exhibited a potent antiviral activity with selectivity index (SI) of more than 22.73. The combined antiherpetic effects of APBP with nucleoside antiherpetic agents, acyclovir (ACV) and vidarabine (ara-A), were examined on the multiplication of these two strains of herpesviruses in Vero cells by the combination assay. The results of combination assay were evaluated by the combination index (CI) that was calculated by the multiple drug effect analysis. CI values were in the range 0.47-0.51 for a combination of APBP with ACV, and in the range of 1.02-1.18 for a combination of APBP with ara-A. The combinations of APBP with ACV on HSV-1 and HSV-2 showed potent synergistic effects, and these results suggest that the possibility of developing APBP as a new antiherpetic agent.

Acyclovir↗

PubMed2000

Antiherpetic activities of acidic protein bound polysacchride isolated from Ganoderma lucidum alone and in combinations with interferons.

To investigate antiherpetic activity, an acidic protein bound polysaccharide (APBP) was isolated from carpophores of Ganoderma lucidum. This brownish APBP was isolated from water soluble substances of the carpophores by activity-guided isolation method. APBP was tested for its antiviral activity against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) by plaque reduction assay in tissue culture. APBP showed potent antiviral activity against HSV-1 and HSV-2 in Vero cells at its 50% effective concentration (EC(50)) of 300 and 440 microg/ml, respectively. APBP had no cytotoxicity on Vero cells at a concentration of 1x10(4) microg/ml. APBP exhibited a potent antiviral activity with selectivity index (SI) of more than 22.73. The combined antiherpetic effects of APBP with protein antiviral agents, interferon alpha (IFN alpha) and interferon gamma (IFN gamma), were examined on the multiplication of these two strains of herpesviruses in Vero cells by the combination assay. The results of combination assay were evaluated by the combination index (CI) that was calculated by the multiple drug effect analysis. The combinations of APBP with IFN alpha on HSV-1 and HSV-2 showed more potent synergistic effects with CI values of 0.30-0.62 for 50-90% effective levels than those of APBP with IFN gamma with CI values of 0.65-1.10. These results suggest the possibility of developing APBP as a new antiherpetic agent.

Animals↗

PubMed2000

Possible mode of antiviral activity of acidic protein bound polysaccharide isolated from Ganoderma lucidum on herpes simplex viruses.

Two protein bound polysaccharides, a neutral protein bound polysaccharide (NPBP) and an acidic protein bound polysaccharide (APBP), were isolated from water soluble substances of Ganoderma lucidum by EtOH precipitation and DEAE-cellulose column chromatography. Their antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were then investigated by plaque reduction assay. APBP exhibited more potent HSV-1 and HSV-2 antiviral activity than NPBP with 50% effective concentration (EC(50)) of 300-520 microg/ml. In order to examine the possible mode of the antiviral activity of APBP its virucidal effect, antiviral activity in preincubation, attachment and penetration assay were tested with HSV-1 and HSV-2. APBP was found to have a direct virucidal effect on HSV-1 and HSV-2. APBP did not induce IFN or IFN-like materials in vitro and is not expected to induce a change from a normal state to an antiviral state. APBP in concentrations of 100 and 90 microg/ml inhibited up to 50% of the attachment of HSV-1 and HSV-2 to Vero cells and was also found to prevent penetration of both types of HSV into Vero cells. These results show that the antiherpetic activity of APBP seems to be related to its binding with HSV-specific glycoproteins responsible for the attachment and penetration, and APBP impedes the complex interactions of viruses with cell plasma membranes.

Animals↗

PubMed1999

Antiviral activities of various water and methanol soluble substances isolated from Ganoderma lucidum.

In order to find antiviral substances from basidiomycetes, two water soluble substances, GLhw and GLlw, and eight methanol soluble substances, GLMe-1-8, were prepared from carpophores of Ganoderma lucidum. These substances were examined for their activities against five strains of pathogenic viruses such as herpes simplex virus types 1 (HSV-1) and 2 (HSV-2), influenza A virus (Flu A) and vesicular stomatitis virus (VSV) Indiana and New Jersey strains in vitro. Antiviral activities were evaluated by the cytopathic effect (CPE) inhibition assay and plaque reduction assay. Five substances, GLhw, GLMe-1, -2, -4 and -7 significantly inhibited the cytopathic effects of HSV and VSV. In the plaque reduction assay, GLhw inhibited plaque formation of HSV-2 with 50% effective concentrations (EC50) of 590 and 580 microg/ml in Vero and HEp-2 cells, and its selectivity indices (SI) were 13.32 and 16.26. GLMe-4 did not exhibit cytotoxicity up to 1000 microg/ml, while it exhibited potent antiviral activity on the VSV New Jersey strain with an SI of more than 5.43. These results indicate the possibility of development of antiviral agents from basidiomycetous fungi.

Animals↗

PubMed1999

Antiherpetic activities of various protein bound polysaccharides isolated from Ganoderma lucidum.

To investigate antiherpetic substances from Ganoderma lucidum, various protein bound polysaccharides, GLhw, GLhw-01, GLhw-02, GLhw-03, were isolated by activity-guided isolation from water soluble substances of the carpophores. These substances were examined for their antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) by plaque reduction assay in vitro. Among them, the acidic protein bound polysaccharide, GLhw-02 of a brownish substance, exhibited the most potent antherpetic activity with 50% effective concentrations (EC50) of 300 approximately 520 microg/ml in Vero and HEp-2 cells, and its selectivity indices (SI) were more than 20. GLhw-02 was identified to consist mainly of polysaccharide (approximately 40.6%) and protein (approximately 7.80%) by anthrone test and Lowry-Folin test, and showed the usual molar ratio (C:H:O = 1:2:1) of carbohydrates by elemental analysis. These results suggest that GLhw-02 possesses the possibility of being developed from a new antiherpetic agent.

Acyclovir↗

PubMed2004

Reusing soy residue for the solid-state fermentation of Ganoderma lucidum.

Lingzhi has been a popular oriental medicine used to treat various human diseases. Soy residue from the waste of tofu manufacturing was used to culture Ganoderma lucidum in solid-state fermentation. The solid-state fermentation was conducted in three types of containers: test tube, 500-ml flask, and sterilize-able polypropylene plastic bag. The highest rate of mycelial growth of 6 mm/day was observed in the medium of carbon/nitrogen (C/N) ratio of 80 using test tubes. However, a growth rate of 7.5 mm/day was found at the C/N ratio of 70-80 in the 500-ml flasks. In the tests using plastic bags, the fruiting bodies were fully developed only for the C/N ratios of 70 and 80. The components of fruiting bodies obtained from different media were also analyzed and compared. The contents of ash, polysaccharides, and crude protein of fruiting bodies were found higher in the media of C/N ratio of 80.

Analysis of Variance↗