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PubMedDate not supplied

[Polysaccharides of Ganoderma lucidum: factors affecting their production].

The conditions of polysaccharide production by the fungus Ganoderma lucidum were optimized. The maximal yield of endopolysaccharides and exopolysaccharides was observed at 25-30 degrees C, initial pH of culture medium 4.0-6.0, and at a C : N ratio of approximately 18 : 1 and 25 : 1, respectively. The greatest yield of mycelium was reached at a more intensive aeration, and the maximal yield of polysaccharides was observed at a less intensive aeration. The optimal ratio between fungus growth and polysaccharide production was observed at 100 rpm and an aeration of 1.0-1.5 1/1 medium min.

Hydrogen-Ion Concentration↗

PubMed2005

[Purification and characteristic of proteinase inhibitor GLPIA2 from Ganoderma lucidum by submerged fermentation].

A proteinase inhibitor GLPIA2 was purified to homogeneity from Ganoderma lucidum by submerged fermentation. The purification was carried out by ethanol fractional precipitation (30%-80%), gel-filtration on Superdex 200 column (30 cm x 1.1 cm i.d.) and anion exchange on Source 30Q column (10 cm x 1.6 cm i.d.). The gel chromatographic conditions were as follows: 50 mmol/L sodium phosphate as mobile phase with a flow rate of 1 mL/min with the effluent collection of 1 mL/tube and detection at 280 nm. The anion exchange chromatographic conditions were as follows: 50 mmol/L Tris-HCl (pH 8.8) containing different amounts of NaCl as mobile phase with a flow rate of 2 mL/min with the effluent collection of 5 mL/tube and detection at both 215 nm and 280 nm. Two active fractions named GLPIA1 and GLPIA2 corresponding to proteinase A inhibitory activities were pooled and lyophilized. GLPIA2 only has the absorption at 215 nm. The relative molecular mass of the inhibitor was 15,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The amino acid composition of GLPIA2 was analyzed by high performance liquid chromatography. The chromatographic conditions were as follows: C18 column (125 mm x 4.0 mm i.d.) with column temperature of 40 degrees C; a mixture of 20 mmol/L sodium acetate-methanol-acetonitrile as mobile phase with a flow rate of 1 mL/min and detection at 338 nm. The results indicate that GLPIA2 is rich in acidic amino acid (Glu) and low in aromatic amino acids (Phe and Tyr). The interaction of some proteinases with GLPIA2 was investigated. The inhibitors are more potent against pepsin and yeast proteinase A than other proteinases.

Chromatography, High Pressure Liquid↗

PubMed2003

[Effects of extracts of Chinese medicines on Ganoderma lucidum in submerged culture].

Effects of water and ethanol extracts of 10 Chinese medicines, such as Astragalus membranaceus, Coix lachryma-jobi, etc., on biomass and exopolysaccharide of Ganoderma lucidum were studied by submerged culture. The results showed: water extracts of all medicines can improve the culture of G. lucidum except of A. membranaceus, ethanol extracts of C. lachryma-jobi, Dioscorea opposita, Codonopsis pilosula, and Achyranthes bidentata( < 187.5g Medicine/L substrate) can also increase the biomass of G. lucidum, but the ethanol extracts of Angelica sinensis, Dendrobium nobile check the growth of G. lucidum. The production of exopolysa-ccharide can be improved by all the Chinese medicines and their dosage used in this experiment, Although A. sinensis, D. Nobile check the growth of G. lucidum, they could stimulate the secretion of exopolysaccharide in lower dosage. It is concluded that some Chinese medicines, such as C. lachryma-jobi, D. opposita, C. pilosula, etc. can be processed by the fermentation of G. lucidum, and bio-active compound can be produced by adding appropriate Chinese medicine in the substrate to culture G. lucidum.

Biomass↗

PubMed2005

[Comparison of the immunomodulatory effects of spore polysaccharides and broken spore polysaccharides isolated from Ganoderma lucidum on murine splenic lymphocytes and peritoneal macrophages in vitro].

OBJECTIVE: To compare the immunomodulatory effects of spore polysaccharides (Gl-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. METHODS: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-gamma and TNF-alpha in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. RESULTS: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-alpha and NO in Gl-SP- or Gl-BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8 mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-gamma in doublejway MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2-12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4(+) or CD8(+) subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4(+) and CD8(+) subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. CONCLUSION: Gl-SP and Gl-BSP have similar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP.

Adjuvants, Immunologic↗

PubMed1991

[In vitro cytotoxicity of Ganoderma lucidum on oral cancer cells].

The extract from the mycelium of Ganoderma lucidum was diluted into serial concentrations and added into in vitro cultured oral cancer and normal cell lines. After incubation for 24 hours, the survival fraction was determined by MTT colorimetric assay. The result revealed that the ID50 was about 3mg/ml and the total lethal dosage was beyond 4 mg/ml. This toxic effect was the same in both cancer and normal cells. Not only was there no difference between cancer and normal cells, but also the high dosage required in toxicity leads to the conclusion that the GL has no direct cytotoxic effect in cancer treatment.

Antineoplastic Agents, Phytogenic↗

PubMedDate not supplied

[Optimization of nutrient medium for submerged cultivation of Ganoderma lucidum (Curt.: Fr.) P. Karst].

The dependence of the amount of the grown vegetative mycelium of Ganoderma lucidum on the composition of the nutrient medium has been studied under conditions of submerged cultivation. The medium was optimized using full factorial and steepest ascent experimental designs. The addition of two carbon sources to the medium considerably improved the submerged growth of the fungus. An optimized medium provided for a high yield (20-20.95 g/l) of the morphologically homogeneous mycelium and shortened the cultivation period to 3-4 days.

Culture Media↗

PubMed2006

Ganoderma lucidum inhibits proliferation of human breast cancer cells by down-regulation of estrogen receptor and NF-kappaB signaling.

Ganoderma lucidum, an oriental medical mushroom, has been used in Asia for the prevention and treatment of a variety of diseases, including cancer. We have previously demonstrated that G. lucidum inhibits growth and induces cell cycle arrest at G0/G1 phase through the inhibition of Akt/NF-kappaB signaling in estrogen-independent human breast cancer cells. However, the molecular mechanism(s) responsible for the inhibitory effects of G. lucidum on the proliferation of estrogen-dependent (MCF-7) and estrogen-independent (MDA-MB-231) breast cancer cells remain to be elucidated. Here, we show that G. lucidum inhibited the proliferation of breast cancer MCF-7 and MDA-MB-231 cells by the modulation of the estrogen receptor (ER) and NF-kappaB signaling. Thus, G. lucidum down-regulated the expression of ERalpha in MCF-7 cells but did not effect the expression of ERbeta in MCF-7 and MDA-MB-231 cells. In addition, G. lucidum inhibited estrogen-dependent as well as constitutive transactivation activity of ER through estrogen response element (ERE) in a reporter gene assay. G. lucidum decreased TNF-alpha-induced (MCF-7) as well as constitutive (MDA-MB-231) activity of NF-kappaB. The inhibition of ER and NF-kappaB pathways resulted in the down-regulation of expression of c-myc, finally suppressing proliferation of estrogen-dependent as well as estrogen-independent cancer cells. Collectively, these results suggest that G. lucidum inhibits proliferation of human breast cancer cells and contain biologically active compounds with specificity against estrogen receptor and NF-kappaB signaling, and implicate G. lucidum as a suitable herb for chemoprevention and chemotherapy of breast cancer.

Breast Neoplasms↗

PubMed2006

Ganoderma lucidum extract stimulates glucose uptake in L6 rat skeletal muscle cells.

The effect of Ganoderma lucidum extract on glucose uptake was studied in L6 rat skeletal muscle cells. G. lucidum extract increased glucose uptake about 2-fold compared to control. The extract stimulated the activity of phosphatidylinositol (PI) 3-kinase which is a major regulatory molecule in the glucose uptake pathway. About 7-fold increased activity of a PI 3-kinase was observed after treatment with G. lucidum extract, whereas PI 3-kinase inhibitor, LY294002, blocked the G. lucidum extract-stimulated PI 3-kinase activity in L6 skeletal muscle cells. Protein kinase B, a downstream mediator of PI 3-kinase, was also activated by G. lucidum extract. We then assessed the activity of AMP-activated protein kinase (AMPK), another regulatory molecule in the glucose uptake pathway. G. lucidum extract increased the phosphorylation level of both AMPK alpha1 and alpha2. Activity of p38 MAPK, a downstream mediator of AMPK, was also increased by G. lucidum extract. Taken together, these results suggest that G. lucidum extract may stimulate glucose uptake, through both PI 3-kinase and AMPK in L6 skeletal muscle cells thereby contributing to glucose homeostasis.

AMP-Activated Protein Kinases↗

PubMed2006

Inhibition of oxidative stress-induced invasiveness of cancer cells by Ganoderma lucidum is mediated through the suppression of interleukin-8 secretion.

Epidemiological studies suggest that the intake of natural/nutrient products is inversely related to cancer risk. While oxidative stress, generating reactive oxygen species, has been linked to cancer initiation and progression, dietary antioxidants have reduced the risk of certain cancers. Experimental studies have demonstrated that antioxidants and phytochemicals could prevent cancer metastasis, and antioxidants were suggested as adjuvants in cancer therapy. Ganoderma lucidum is an Asian medicinal mushroom that has been used for the past two thousand years for the treatment of various diseases, including cancer. G. lucidum is currently popular as a dietary supplement in the form of tea, powder or extract. We have previously demonstrated that G. lucidum suppresses growth, angiogenesis and invasiveness of highly invasive and metastatic breast cancer cells. The present study was undertaken to evaluate the effect of G. lucidum on oxidative stress-induced metastatic behavior of poorly-invasive MCF-7 breast cancer cells. We show that G. lucidum inhibits oxidative stress-induced migration of MCF-7 cells by the down-regulation of MAPK signaling. G. lucidum suppressed oxidative stress stimulated phosphorylation of extracellular signal-regulated protein kinases (Erk1/2), which resulted in the down-regulation of expression of c-fos, and in the inhibition of transcription factors AP-1 and NF-kappaB. The biological effect of G. lucidum on cell migration was mediated by the suppression of secretion of interleukin-8 from MCF-7 cells exposed to oxidative stress. In summary, our results suggest that G. lucidum inhibits the oxidative stress-induced invasive behavior of breast cancer cells by modulating Erk1/2 signaling and can be potentially considered as an antioxidant in adjuvant cancer therapy.

Antioxidants↗

PubMed2006

Inhibitory effect of a water-soluble extract from the culture medium of Ganoderma lucidum (Rei-shi) mycelia on the development of pulmonary adenocarcinoma induced by N-nitrosobis (2-hydroxypropyl) amine in Wistar rats.

A water-soluble extract from the culture medium of Ganoderma lucidum (Rei-shi) mycelia (MAK) has been shown to exert a potent chemopreventive effect. The present study was designed to investigate the effects of dietary MAK supplementation on the development of lung tumors initiated by N-nitrosobis (2-hydroxypropyl) amine (BHP) in male Slc:Wistar rats. A total of 77 animals, 6 weeks of age, were divided into 5 groups and given BHP (2,000 ppm) in their drinking water for 10 weeks. The normal controls were not supplied with BHP. After treatment with the carcinogen, the rats were fed a normal control MF solid diet, or the same diet containing MAK (1.25%, 2.5% or 5%) for 12 weeks. Macroscopically, all the doses of MAK reduced the number of nodules, and the effect of 5% MAK was found to be especially significant. Microscopically, an increase in the number of proliferating cell nuclear antigen (PCNA)-negative tumors and a decrease in the number of tumors strongly positive for PCNA were observed in the tissue sections from the rats that had received all the doses of MAK. The present results thus indicate that dietary supplementation with MAK inhibits the development of lung tumors, suggesting that MAK may be a potent chemopreventive agent against lung carcinogenesis.

Adenocarcinoma↗

PubMed2006

Antiproliferative ability of a combination regimen of crocodile egg extract, wild radix ginseng and natural Ganoderma lucidum on acute myelogenous leukemia.

Chinese practitioners have employed the use of traditional Chinese medicine as an anti-cancer agent since the ancient period. Different combinations have been formulated for various purposes. Some have been claimed for post-chemotherapy use but their direct actions on cancer cells may not be significantly reported. In the present study, we have tested the possible anti-leukemia potential of a combination regimen including crocodile egg extract, wild radix ginseng and natural Ganoderma lucidum (CGG extract) on acute myelogenous leukemia (AML) in vitro. A water soluble CGG extract was prepared and its antiproliferative activity was tested on the KG1a AML cell line and two freshly prepared bone marrow aspirate samples isolated from patients with de novo AML during presentation by a MTS/PMS assay. Furthermore, the possible activity of the CGG extract on the regeneration potential of KG1a cells was also investigated using a semi-solid methyl-cellulose colony formation assay. Lastly, the acute toxicity of CGG extract was further examined by a single high-dose oral feeding to rats. We found that the CGG extract could possess significant antiproliferative activity on AML cells. A strong colony formation inhibition was further demonstrated on KG1a cells. After feeding the rats with an excessive dose of CGG extract, we observed no development of acute toxicity. We concluded that the CGG extract has growth inhibitory potential on KG1a cells and AML bone marrow samples in vitro. An in vivo toxicity test revealed that no acute toxicity was observed after feeding the rats a high dosage of the CGG extract. Further animal model tests are necessary to investigate the possible chronic toxicity of the CGG extract.

Alligators and Crocodiles↗

PubMed1991

Molecular cloning of a cDNA and a gene encoding an immunomodulatory protein, Ling Zhi-8, from a fungus, Ganoderma lucidum.

A large amount of the novel immunomodulatory protein Ling Zhi-8 (LZ-8) is synthesized in the mycelia of Ganoderma lucidum (Kino, K., Yamashita, A., Yamaoka, K., Watanabe, J., Tanaka, S., Ko, K., Shimizu, K., and Tsunoo, H. (1989) J. Biol. Chem. 264, 472-478). A cDNA and a gene for LZ-8 were isolated and characterized. The mixed oligonucleotide probes for LZ-8 cDNA were designed from the results of protein sequencing (Tanaka, S., Ko, K., Kino, K., Tsuchiya, K., Yamashita, A., Murasugi, A., Sakuma, S., and Tsunoo, H. (1989) J. Biol. Chem. 264, 16372-16377) and were used for screening the mycelial cDNA library. The nucleotide sequence of the cloned cDNA confirms the amino acid sequence of LZ-8 that was previously determined by protein sequencing. The clones containing the LZ-8 gene (lz-8) were obtained from the mycelial genomic DNA library using the cDNA probe. Two CCAAT-like sequences and one TATA box were found at the upstream region of the postulated transcription initiation site of lz-8. A small intron (61 nucleotides long) divided lz-8 into two exons at the 5'-untranslated region. The other characteristic sequences were also found around the postulated transcription initiation site and around the poly(A) additional site.

Adjuvants, Immunologic↗

PubMed1990

[Studies on constituents of ganoderma capense IV. The chemical structures of ganoine, ganodine and ganoderpurine].

Two novel pyrrole alkaloids--ganoine and ganodine and a novel purine alkaloid ganoderpurine have been isolated from the mycelium of Ganoderma capense (Lloyd)Teng (Polyporaceae) obtained by submerged fermentation. On the basis of spectroscopic data their structures were elucidated, ganoine is N-isopentyl-5-hydroxymethyl-pyrryl alldehyde. Ganodine is N-phenylethyl-5-hydroxymethyl-pyrryl aldehyde and ganoderpurine is N9-(alpha, alpha dimethyl-gamma-oxobutyl) adenine.

Adenine↗

PubMed1989

Complete amino acid sequence of an immunomodulatory protein, ling zhi-8 (LZ-8). An immunomodulator from a fungus, Ganoderma lucidium, having similarity to immunoglobulin variable regions.

The complete amino acid sequence of a novel immunomodulatory protein, ling zhi-8 (LZ-8), isolated from a fungus, Ganoderma lucidium (Kino, K., Yamashita, A., Yamaoka, K., Watanabe, J., Tanaka, S., Ko, K., Shimizu, K., and Tsunoo, H. (1989) J. Biol. Chem. 264, 472-478), was determined by protein sequencing. The polypeptide consists of 110 amino acid residues with an acetylated amino end and has a molecular mass of 12,420 Da including an amino-end blocking group. There is no attachment site for an Asn-linked oligosaccharide chain, consistent with the very low carbohydrate content of LZ-8. These results indicate that the native form of LZ-8 with a molecular mass of 24 kDa is a homodimer of the LZ-8 polypeptide whose sequence is described here. Furthermore, the LZ-8 chain shows considerable similarity to the variable region of immunoglobulin heavy chain both in its sequence and in its predicted secondary structure. The interesting possibility that LZ-8 is related to an ancestral protein of the immunoglobulin superfamily is also discussed.

Adjuvants, Immunologic↗

PubMed1989

Isolation and characterization of a new immunomodulatory protein, ling zhi-8 (LZ-8), from Ganoderma lucidium.

A novel protein with mitogenic activity in vitro and immunomodulating activity in vivo has been isolated from the mycelial extract of an Oriental medicinal fungus, ling zhi (Ganoderma lucidium). This protein was named ling zhi-8 (LZ-8) and its biochemical and immunological properties are described. LZ-8 was purified by two chromatographic systems, gel filtration and followed by ion-exchange, using an in vitro bioassay measuring blast-formation stimulatory activity toward mouse spleen lymphocytes to monitor purification. Analysis by several types of electrophoresis revealed a single band, with the molecular weight differing slightly depending on the system employed. Under reduced conditions, sodium dodecyl sulfate-polyacrylamide gel electrophoresis using the method of Laemmli, U.K. ((1970) Nature 227, 680-685) indicated an apparent Mr = 17,100, while under nonreduced conditions an apparent Mr = 17,500 was found; and, using Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a value of apparent Mr = 13,100 was obtained. LZ-8 has an isoelectric point of 4.4, and sugar analysis indicated a low carbohydrate content (1.3%). Half-cysteine, histidine, and methionine were not detected from the analysis of amino acid composition after further purification of LZ-8 by reversed-phase high performance liquid chromatography. LZ-8 was capable of hemagglutinating sheep red blood cells, but no such activity was observed toward human red blood cells (A, B, AB, and O types). In vivo, LZ-8 prevents the production of systemic anaphylaxis reaction in mice if it has been administered repeatedly, and reduction of antibody production is the suggested mechanism. The mechanisms of hemagglutination of sheep red blood cells and of blast-formation stimulation of mouse spleen cells are also discussed.

Adjuvants, Immunologic↗

PubMed1995

[Psychoneuroimmunological effects of morphine and the immunoprotection of Ganoderma polysaccharides peptide in morphine-dependent mice].

Different kinds of single, multiple, acute or chronic administrations of morphinized animal models were established, with which a series of experiments in both in vivo and in vitro systems as well as molecular levels were pharmacologically designed to investigate the psychoneuroimmunological effects of morphine (Mor) and the immunoprotection of Ganoderma polysaccharides peptide (GPP) in Mor-dependent mice. It was first discovered that both c-myb and c-myc mRNA expression in splenocytes of repetitive Mor-treated mice were detected to be significantly decreased, and that GPP could induce restoration of several immunologic parameters depressed by Mor treatment to or even beyond normal levels. This provides the experimental animal evidence that immune response modifiers such as GPP could be of potential application in controlling abuse of opiates-induced immunodeficiency.

Animals↗

PubMed1993

[New techniques of cultivating Ganoderma lucidum (W. Curt.:Fr)Karst.,Rev. with woodlog].

Imitating wild cultivation of Ganoderma lucidum with short-woodlog is a method of cultivating artificial G. lucidum developed in recent years. The method can be applied to large scale production. This paper shows that section-inoculating and impregnating with steamed shortwoodlog indoors and soil-cover cultivating under large shed are important in the cultivation.

Methods↗

PubMed1993

[Effects of Ganoderma polysaccharides on the activity of DNA polymerase alpha of splenocytes and immune function in aged mice].

The activity of DNA polymerase alpha in splenocytes of 24-month-old mice was about 35.6% lower than that of 3-month-old mice. Aged mice were intraperitoneally administered Ganoderma polysaccharides (GL-B) once a day for 4 days and then the activity of the enzyme was assessed. The results showed that GL-B at doses of 25 and 50 mg/kg-1 enhanced the activity of the enzyme in aged mouse splenocytes by 44.0 and 58.8% respectively. In addition, the mixed lymphocyte response to alloantigen, automatic proliferation and IL-2 production of splenocytes in aged mice declined as compared with that in young adult mice. GL-B (50, 100, 200 micrograms.ml-1) was found to restore those parameters to the levels of that of young mice in vitro.

Aging↗