Structural characterization and anti-oxidative activity for a glycopeptide from Ganoderma lucidum fruiting body.

A water-soluble glycopeptide (named GL-PWQ3) with a molecular weight (Mw) of 2.40 × 104 g/mol was isolated from Ganoderma lucidum fruiting body by hot water extraction, membrane ultrafiltration, and gel column chromatography, which mainly consisted of glucose and galactose. Based on the methylation, FT-IR, 1D, and 2D NMR analysis, the polysaccharide portion of GL-PWQ3 was identified as a glucogalactan, which was comprised of unsubstituted (1,6-α-Galp, 1,6-ß-Glcp, 1,4-ß-Glcp) and monosubstituted (1,2,6-α-Galp and 1,3,6-ß-Glcp) in the backbone and possible branches that at the O-3 position of 1,3-Glcp and T-Glcp, and the O-2 position of T-Fucp, T-Manp or T-Glcp. The chain conformational study by SEC-MALLS-RI and AFM revealed that GL-PWQ3 was identified as a highly branched polysaccharide with a polydispersity index of 1.25, and might have compact sphere structures caused by stacked multiple chains. Moreover, the GL-PWQ3 shows strong anti-oxidative activity in NRK-52E cells. This study provides a theoretical basis for further elucidating the structure-functionality relationships of GL-PWQ3 and its potential application as a natural antioxidant in pharmacotherapy as well as functional food additives.

[Effect of Ganoderma lucidum polysaccharides peptide on invasion of human lung carcinoma cells in vitro].

OBJECTIVE: To find out the effects of Ganoderma lucidum polysaccharides peptide (Gl-PP) on the invasion of the human lung carcinoma cell (PG cell). METHODS: PG cells were pretreated with different concentration Gl-PP in vitro, using cell proliferation assay, cell migration assay, adhesion assay, zymography and RT-PCR, then the effects of Gl-PP on proliferation, motility, adhesion and MMP-9 activity and mRNA expression of PG cells were investigated in vitro. RESULTS: Gl-PP did not directly inhibit PG cell proliferation in vitro. However, pretreated with Gl-PP, PG cells motility was inhibited significantly. PG cells adhesion was also inhibited. The activity of MMP-9 was inhibited in a dose-dependent manner, and the inhibited ratio was 41.53% at a dose of 100 mg/L Gl-PP. The mRNA expression of MMP-9 of pretreated PG cells was inhibited. CONCLUSION: Gl-PP could suppress invasion of human lung carcinoma cells in vitro.

[Isolation, purification and structural analysis of GL-PP-3A, an active polysaccharide peptide from Ganoderma lucidum].

GL-PP-3A, an active polysaccharide peptide, was isolated and purified from Ganoderma lucidum, and then its structure was analyzed. Crude polysaccharide peptides were extracted from Ganoderma lucidum with hot water, precipitated with ethanol and then dialyzed from Ganoderma lucidum. Subsequently GL-PP-3A was isolated and purified from the crude polysaccharide peptides by fractional precipitation and chromatography of Bio-Gel P-10 column. The repetitive unit of GL-PP-3A was analyzed by high performance gel permeation chromatography (HPGPC), monosaccharide composition and methylation analysis, 1H NMR and 13C NMR. GL-PP-3A is a heteropolysaccharide which is composed mainly of glucose (Glc), and also contains saccharide residues such as rhamnose (Rha), xylose (Xyl), mannose (Man) and galactose (Gal) and 17 kinds of amino acids. Its weight-average molecular weight (Mw) and number-average molecular weight (Mn) were 1.7 x 10(4) and 1.1 x 10(4), respectively, with the ratio of Mw/Mn ( molecular weight distribution) being of 1.49. Its backbone chain is composed of 1,6-linked beta-D-Glcp and 1,3-liked beta-D-Glcp at a ratio of 2:1. Some of 1,6-linked glucose residuals of the backbone chain are substituted at 2-0 or 3-0, and there are 1 to 3 1,6-linked beta-D-Galp or 1,3-linked alpha-D-Manp in the branched chains, the nonreducing ends of which consist mainly of beta-D-Glcp and a few Rha.

[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.

Ganoderma total sterol (GS) and GS1 protect rat cerebral cortical neurons from hypoxia/reoxygenation injury.

The effect of Ganoderma total sterol (GS) and its main components(GS(1)) on rat cortical neuronal cultures exposed to hypoxia/reoxygenation (H/R) was studied in vitro. GS (0.01,0.1,1 microg/ml) increased neuron viability following H/R. GS also significantly reduced malondialdehyde content and reactive oxygen species production and increased manganese superoxide dismutase (Mn-SOD) activity; furthermore, the translocation of nuclear factor-kappa B and the production of interleukin-1beta and tumor necrosis factor alpha induced by H/R were also blocked. These findings suggest that GS might be useful in treating H/R-induced oxidative stress and inflammatory response. We also hypothesized that Mn-SOD might play a critical role in the neuroprotective effect of GS against H/R injury. In addition, pretreatment with GS(1) (0.01, 0.1, 1 microg/ml) significantly attenuated the decline of neuron viability and the formation of reactive oxygen species. Furthermore GS(1) possessed more potent protective effect on neurons compared with GS at the same dose. These findings demonstrated that GS(1) is the main component in GS; and play a critical role in the neuroprotective effect of GS against H/R.