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Ganoderma lucidum: partial characterization of spore and whole body antigenic extracts.

This study focused on the characterization of antigenic/ allergenic profiles of Ganoderma lucidum spore and whole body preparations. Whole body G. lucidum contained higher protein to carbohydrate ratio whereas it was less than one for spore extract. Isoelectric focusing showed 12 and 11 bands in acidic pH range (pI 3.5-6.5) for G. lucidum spore and whole body, respectively, while SDS-PAGE showed 8 and 23 fractions, respectively, in molecular weight range of 12.8-75.0 kD. The prominent protein fractions of G. lucidum spores were 19.4, 22.8 and 23.8 kD, whereas for G. lucidum whole body, 13.2, 14.7, 18. 7, 21.5 and 23.5 constituted major fractions. Immunoblotting with 41 individual serum samples revealed 21.8, 23.8, 19.4 and 20.0 kD to be major allergenic protein fractions of G. lucidum spores. The same using G. lucidum whole body and 26 individual serum samples identified several fractions of 17.0, 17.5, 18.5, 22.0, 23.8, 42.0, 44.0, 56.0 and 69.0 kD as major allergens. The compiled data suggest that there are common as well as specific allergenic components in two G. lucidum extracts studied.

Allergens↗

[Chemical studies on peptidepolysaccharides of Ganoderma lucidum (W. Curt. Fr.) Karst].

Seven homogeneous peptidepolysaccharides have been obtained from the fruit-body of Ganoderma lucidum. The main constituents are TGLP-2, TGLP-3, TGLP-6 and TGLP-7, whose molecular weights are 20.9 x 10(4), 4.5 x 10(4), 3.2 x 10(4) and 10 x 10(4) respectively. TGLP-2 is a heteroglycan peptide, TGLP-3 and TGLP-6 are glycan peptides and TGLP-7 is a galactan peptide.

Drugs, Chinese Herbal↗

[Studies on the ganoderic acid, a new constituents from the fruiting body of Ganoderma lucidum (Fr.) Karst].

Three compounds have been isolated from the dichloromethane soluble fraction of the fruiting body of Ganoderma lucidum (Fr.) Karst. On basis of spectral analyses (UV, IR, MS, 1HNMR, 13CNMR and 2D-NMR), they were identified as 3, 7-dioxo-lanosta-8, 24(E)-dien-26-oic acid (I), 7 beta-15 alpha-dihydroxy-3, 11, 23-trioxo-5 alpha-lanost-8-en-26-oic acid (II) and 3 beta, 7 beta, 15 alpha-trihydroxy-11, 23-dioxo-5 alpha-lanosta-8-en-26-oic acid (III). Compound I is a new compound named ganoderic acid DM.

Drugs, Chinese Herbal↗

Effects of polysaccharides (FI0-b) from mycelium of Ganoderma tsugae on proinflammatory cytokine production by THP-1 cells and human PBMC (I).

AIM: To compare the effects of water-soluble polysaccharides, FI0-b, and its formic acid-modified derivative, FI0-b-H, on production of human proinflammatory cytokines. METHODS: The polysaccharides were modified by formic acid. Cytokine production was quantitated by radioimmunoassay. mRNA for cytokines was measured by semi-quantitative RT-PCR. RESULTS: FI0-b and FI0-b-H 4, 40, and 400 mg/L significantly downregulated interleukin-1 alpha (IL-1 alpha) production by THP-1 cells induced by lypopolysaccharide (LPS) 1 or 10 mg/L and phorbol myristate acetate (PMA) 200 nmol/L. At lower stimulation with LPS 10 mg/L and PMA 200 nmol/L, both polysaccharides significantly upregulated tumor necrosis factor alpha (TNF alpha) production by THP-1 cells. However, at higher stimulation with LPS 100 mg/L and PMA 200 nmol/L, they downregulated TNF alpha production. FI0-b-H downregulated interleukin-8 (IL-8) production by THP-1 cells at a lower-dose of LPS 1 mg/L and PMA 200 nmol/L, but upregulated IL-8 production at a higher-dose of LPS 10 mg/L and PMA 200 nmol/L. Production of cytokines (IL-1 alpha and TNF alpha) was transcriptionally or post-transcriptionally regulated by FI0-b and FI0-b-H. CONCLUSION: The water-soluble polysaccharides of Ganoderma tsugae mycelium have bidirectional immunomodulatory effects on cytokine production in different stimulatory conditions in a dose-dependent manner. Compared with FI0-b, FI0-b-H has more marked effects on human proinflammatory cytokine production.

Adult↗

Effects of polysaccharides (FI0-c) from mycelium of Ganoderma tsugae on proinflammatory cytokine production by THP-1 cells and human PBMC (II).

AIM: To study the effects of water-soluble polysaccharides. FI0-c, and its sulfated derivative, FI0-c-S, on production of human proinflammatory cytokines, interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor alpha (TNF alpha). METHODS: The herbal polysaccharides were modified by chlorosulfornic acid in dimethyl sulfoxide (Me2SO). Cytokine production was measured by radioimmunoassay, mRNA for the cytokines was measured by semi-quantitative RT-PCR. RESULTS: FI0-c 4 mg/L itself induced IL-1 alpha production by THP-1 cells without stimulants, such as lipopolysaccharides (LPS) and phorbol myristate acetate (PMA). On the other hand, FI0-c and FI0-c-S inhibited the IL-1 alpha production by THP-1 cells with these stimulants. FI0-c and FI0-c-S significantly upregulated TNF alpha production by THP-1 cells without stimulants or at a low dose of LPS 10 mg/L and PMA 200 nmol/L, whereas these polysaccharides markedly downregulated the TNF alpha production by a high dose of LPS 100 mg/L and PMA. Human peripheral blood mononuclear cells (PBMC) responded to FI0-c and FI0-c-S in IL-1 alpha and TNF alpha production in a fashion similar to THP-1 cell responses. FI0-c 4 mg/L downregulated high-dose LPS- and PMA-induced IL-1 alpha or TNF alpha mRNA and their protein production by THP-1 cells. CONCLUSION: The water-soluble polysaccharides of Ganoderma tsugae mycelium have bidirectional immunomodulatory effects on cytokine production in different cell stimulatory conditions. Chemical modification of this polysaccharide changed the intensity of regulatory effect on cytokine production.

Adult↗

[Study on antitumor activity and mechanism of Ganoderma polysaccharides B].

OBJECTIVE: To study the antitumor activity and mechanism of Ganoderma polysaccharides (GL-B). METHODS: Both in vivo and in vitro experiments were conducted in the study. Proliferation of tumor cells was detected by MTT method, tumor necrosis factor alpha (TNF alpha) was detected by biological assay, and interferon gamma (IFN gamma) by enzyme-linked immunosorbent assay (ELISA). The level of mRNA was detected with the method of reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: (1) GL-B 50, 100, 200 micrograms/ml inhibited the growth of implanted sarcoma 180 in vivo significantly and dose-dependently. (2) GL-B directly adding to the cultured medium neither induced HL-60 apoptosis nor restrained its proliferation in vitro. (3) The macrophages and T lymphocytes cultured medium treated with GL-B significantly induced HL-60 apoptosis and inhibited its proliferation. GL-B significantly increased TNF alpha and IFN gamma release and their mRNA expression in the cultured medium. CONCLUSION: The anti-tumor activity of GL-B was derived from promoting mRNA expression of TNF alpha and IFN gamma, resulting in TNF alpha and IFN gamma release.

Animals↗

Prevention of development of N,N'-dimethylhydrazine-induced colon tumors by a water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi) mycelia in male ICR mice.

The protective effects of a dietary water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi or Mannentake) mycelia (designated as MAK) against development of colon tumors were investigated in male ICR mice. The animals were given weekly injections of N,N'-dimethylhydrazine (DMH, 10 mg/kg body weight) for the initial 10 weeks to induce colon carcinogenesis, and then fed on diet with or without 5% MAK for 10 weeks. There were no significant differences in incidence and the total number of colon tumors between the groups. However, the MAK diet group demonstrated significantly reduced sizes of tumors in comparison with the MF diet group. Moreover, this was linked to a lowered PCNA positive index and shortening of the germinal region in the colon. beta-catenin positive tumor cell nuclei were also significantly decreased in the MAK group. The present results thus indicate that dietary MAK could act as a potent chemopreventive agent for colon carcinogenesis.

1,2-Dimethylhydrazine↗

Structure and Conformation Behavior of a Glucan from Spores of Ganoderma lucidum (Fr.) Karst.

A novel polysaccharide designated as beta-D-glucan LB-NB, [alpha](D)(21) -24.52(0)(c =0.81, H(2)O)and Mr 4.7x10(4), was isolated and purified from a 0.5 mol/L sodium hydroxide extract of the sporoderm-broken spores of Ganoderma lucidum. The results of NMR experiments, total hydrolysis, methylation analysis and Smith degradation of LB-NB showed that it possesses a backbone consisting of(1right curved arrow 3)- beta-D-glucosyl residues, about two out of nine glucosyl residues being substituted at O-6 by single glucosyl groups. The conformational behavior of LB-NB was studied. Measurements by specific rotations and viscometry showed that the glucan LB-NB takes an ordered structure in water or lower concentration(<0.05 mol/L)alkaline solution and dissolves as random coil in higher concentration(>0.1 mol/L)alkaline solution or in dimethyl sulfoxide(Me(2)SO). The ordered conformation melts into flexible chains with the increase of the concentration of alkali or the V(DMSO)(v/v)of H(2)O-Me(2)SO mixture. The former dissociation is reversible and the later is irreversible. The complex-formation with Congo red indicated that the LB-NB takes neither triple-stranded helical nor randomly coiled conformation in neutral or slightly alkaline solution, and most probably, it should contain single helical structure in aqueous solution. According to immunological test, the LB-NB showed remarkable activity of stimulating the proliferation of T-Cells in vitro, giving a good example for Kulicke's statement.

Journal Article↗

[Effects of Ganoderma lucidum (Leyss ex Fr) Karst compound on the proliferation and differentiation of K562 leukemic cells].

A series of experiments including cell culture and benzidine staining test were undertaken to investigate the effects of Ganoderma lucidum(Leyss ex Fr) Karst Compound(GLC) on the proliferation and differentiation of K562 leukemic cells. The results showed that different concentrations of GLC(from 4 mg.ml-1 to 12 mg.ml-1) could promote human bone marrow granulocyte-macrophage colony forming unit (CFU-GM) proliferation, but suppressed the growth of K562 leukemic cell colonies, and IC50 was 9.2 mg.ml-1. The data from liquid culture demonstrated that GLC could suppress K562 cells proliferation in a dose-dependent(from 4 mg.ml-1 to 20 mg.ml-1) and time-dependent(from 1-5 days) manner. K562 cells could be induced to differentiate into more mature erythrocytic cells by 4 mg.ml-1 and 8 mg.ml-1 GLC. It is concluded that GLC may be a good medicine for leukemia therapy.

Cell Differentiation↗

Polysaccharide purified from Ganoderma lucidum inhibits spontaneous and Fas-mediated apoptosis in human neutrophils through activation of the phosphatidylinositol 3 kinase/Akt signaling pathway.

Ganoderma lucidum has been widely used as a remedy to promote health and longevity in China. The polysaccharide component with a branched (1-->3)-beta-D-glucan moiety from G. lucidum (PS-G) has shown evidence of enhancement of immune responses and of eliciting anti-tumor effects. In this study, we investigated the effect of PS-G on neutrophil viability, which is manifested by spontaneous apoptosis. Annexin V staining and MTT assays reveal that PS-G is able to inhibit spontaneous and Fas-induced neutrophil apoptosis, and this effect of PS-G is enhanced by the presence of zVAD (a caspase inhibitor) and GM-CSF. The antiapoptotic effect of PS-G is diminished by the presence of wortmannin and LY294002 (two PI-3K inhibitors), but is not altered by PD98059 (a MEK inhibitor). Western blotting indicates the stimulating effect of PS-G on Akt phosphorylation and its inhibition of procaspase 3 degradation, which occurs in neutrophils undergoing spontaneous apoptosis or triggered death by Fas. Taken together, PS-G elicitation of antiapoptotic effects on neutrophils primarily relies on activation of Akt-regulated signaling pathways.

Antioxidants↗

Protective effects of Ganoderma lucidum polysaccharides peptide on injury of macrophages induced by reactive oxygen species.

AIM: To study the protective effects of Ganoderma lucidum polysaccharides peptide (GLPP) on the mice peritoneal macrophages injured by reactive oxygen species (ROS), derived from tert-butylhydroperoxide (tBOOH) in vitro and in vivo. METHODS: Mice peritoneal macrophages were injured by ROS, derived from tBOOH. The survival rate of macrophages was measured by MTT assay, and the morphological changes of macrophages were observed under light and electron microscopes. RESULTS: GLPP (50, 100, 200 mg/kg, ip for 5 d) could inhibit the foam cell formation and necrosis of macrophages. The survival rate of macrophages was increased. GLPP (3.125, 12.5, 50, 200 mg/L) given to the cultured macrophages brought the same protective effects. Under the electron microscope it was found that GLPP (100 mg/kg, ip, for 5 d) could protect the organelle such as mitochondria against injury by tBOOH. CONCLUSION: GLPP had significant scavenging ROS and antioxidant effects.

Animals↗

[Extraction and separation of antitumor components from Ganoderma lucidum (Leyss. ex Fr.) Karst].

OBJECTIVE: To examine the variation of alcohol extraction rates of Ganoderma lucidum spores with different rates of wall-wrack, and analyze the antitumor components of alcohol extract by chromatography. METHOD: The G. lucidum spores were soaked and extracted with absolute alcohol. The alcohol extract was chromatographed on a silica gel column and HPLC in proper order, and the antitumor activity of every eluted fraction was represented by its cytotoxicity towards Hela cells. RESULT: Extraction rates 5%, 25% and 33% corresponded to wall-wrack rates 0%, 60%-80% and 99% respectively. The alcohol extract from spores with the highest wall-wrack rate was chromatographed on a silica gel column, eluting successfully with CHCl3, EtOAc and CH3OH in order. The CHCl3 fraction had not any antitumor activity, while this activity of CH3OH fraction was 34 times greater than that of EtOAc fraction. HPLC analysis found out that two mixtures(II1 and II3) possess significant antitumor activity in vitro. CONCLUSION: The weight of alcohol extract from spores with wall-wrack was far greater than that of spores without. The antitumor components of G. lucidum spores could be analyzed with methanol-water on a reverse HPLC.

Antineoplastic Agents, Phytogenic↗

[Studies on difference between sporoderm-broken and nonbroken spores of Ganoderma lucidum (Leyss. ex Fr.) Karst. by polysaccharide analysis].

OBJECTIVE: To compare the release ability of water-soluble polysaccharides in sporoderm-broken and nonbroken spores of Ganoderma lucidum, and establish a comparatively correct method for the determination and analysis of polysaccharide contents in Chinese herbs. METHOD: The release ability of water-soluble polysaccharides was determined on the basis of phenol-sulfuric acid modification in different conditions. RESULT: The release ability of polysaccharides of sporoderm-broken spores was much greater than that of nonbroken spores; and the phenol-sulfuric acid modified cation method proved excellent in accuracy and reproducibility, with a relative error less than 1.5%. CONCLUSION: The spores should be wall-wracked if used as a nutriment, or for extraction and analysis of their effective components. The method can be successfully used for the determination of polysaccharide contents in Chinese herbs or nutriments.

Polysaccharides↗

[Isolation, purification and bioactivities of exopoly saccharides from fermented broth of Ganoderma lucidum].

The exopolysaccharides of Ganoderma lucidum(GLEP) extracted from the fermentation broth after removing protein by Sevage and protease digestion procedures, were applied to a column of DEAE-cellulose(OH- form), and eluted stepwise with distilled water, sodium hydrogen carbonate (0.1 mol/L, 0.3 mol/L, 0.5 mol/L successively) and 0.1 mol/L sodium hydroxide. Five fractions were obtained, and the main fraction was known as GLEP-I, furthermore subjected to chromatography on a column of SepharoseC1-6B, eluted at a flow rate of 30 mL/(cm2.h), the relative viscosity of sample solution of 1.5. Two fractions, GLEP-IFr1 and GLEP-IFr2 with a ratio of 3.8:1, were obtained. Molecular weight of GLEP-IFr1 and GLEP-IFr2 was estimated to be 38,000 and 22,000 Dalton respectively by Membrane Osmometer. The animal test showed that GLEP-IFr1 could inhibited the growth of Sarcoma 180 tumor in mice. The average inhibition ratio was 57.4% (i.p. 10 mg/kg for 10 days). The result of immunological activity showed that GLEP-IFr1 could significantly improve macrophage cytophagy.

Animals↗

[The study on the fermentation medium of Ganoderma lucidum].

This study has explored the fermentation medium of Ganoderma lucidum including inorganic salts, carbon source, nitrogen source and vitamins. Then a suitable formula of fermented medium was obtained by comparing different production slats of Gandoerma powder during fermentation.

Carbon↗

A water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi) mycelia suppresses azoxymethane-induction of colon cancers in male F344 rats.

The present study was designed to investigate the protective effect of a dietary water-soluble extract from cultured medium of Ganoderma lucidum (Rei-shi or Mannentake) mycelia (designated as MAK) on the induction and development of azoxymethane (AOM)-induced colon tumors in male F344/Du Crj rats. A total of 80 animals were divided into five groups at six weeks of age, groups 2, 3 and 4 being given weekly subcutaneous injections of AOM (15 mg/kg body weight) for the initial 3 weeks to induce colon tumors. Rats in group 1 and 5 were injected with the vehicle, 0.9% (w/v) saline, following the same schedule. Rats in groups 1, 2, 3, 4 and 5 were fed MF, MF, 1.25% MAK, 2.5% MAK and 2.5% MAK diets, respectively, starting 1 week before AOM treatment and throughout the six-month experimental period. There were no significant differences in number of ACF, total AC and AC per site among groups 2 to 4, but the tumor incidence was significantly lower, and tumor size was smaller in group 4 (AOM + 2.5% MAK) than in group 2 (AOM + MF). Additionally, beta-catenin positive tumor cell nuclei were significantly decreased in the MAK-fed rats (groups 3 and 4), which also demonstrated lowering of the PCNA labeling index and a shortened germinal region in the colon. The present results thus indicate that dietary MAK could act as a potent chemopreventive agent for colon carcinogenesis.

Adenoma↗