| Phellinus linteus as a rare and precious medicinal fungus, its outstanding physiological effects prepared so concerned, especially the effects of anti-tumor. Mainly research to Phellinus fruiting bodies polysaccharides at present, but the fruiting bodies are scarce and solid-state fermentation of mycelium polysaccharides has been few reported. The research was about the mycelium polysaccharides extraction and purification which was produced by the solid-state fermentation of my laboratory, and the structures, the physiologically activities, the physical and chemical properties. The study provided theoretical basis for developing of health care large-scale products.Mycelium of Phellinus linteus were dealt with crushing, sieving,90℃water bath for3h, removing starch and protein, and precipitated with ethanol, using DEAE-52, G100separation in turn, got three-component polysaccharides finally. The results of all band ultraviolet scanning and the HPLC of three kinds of Phellinus linteus polysaccharides showed that they were all homogeneous polysaccharides named Plps-1, Plps-2, and Plps-3.The relative molecular weight could be measured by gel-chromatography; Plps-1is calculated as2.5×105Da, Plps-2as1.25×105Da, and Plps-3as2.8×104Da. We knew that Plps-1could contain fucose:arabia:sugar:xylose:glucose: galactose=1.336:1.182:1:21.964:1by GC analysis the monosaccharide composition; The Plps-2contains Arabia sugar:fucose:xylose glucose:galactose: mannose:=3.780:1.912:1:3.372:10.430:3.898; The Plps-3contains arabia sugar:fucose: xylose glucose:galactose:mannose:=1.552:1.466:1:1.956:14.368:2.594. Glycosyl GC analysis showed that Plps-1glycosylated1â†'4bonding ratio was63.97%, using glycosylated1â†'3bonding ratio was12.61%, using glycosylated1â†'6bonding or non-reducing end glycosyl ratio was23.42%; glycosylation in Plps-21â†'>4bonding ratio was61.09%,using glycosylated1â†'3bonding ratio was14.05%,using glycosylated1â†'6bonding or non-reducing end glycosyl ratio was24.86%; glycosylation in Plps-31â†'4bonding ratio was11.71%,to glycosyl1â†'3bonding ratio was46.85%,to1â†'6bonding glycosyl or non-reducing end glycosyl ratio for41.44%. According to the infrared spectra could be seen in Plps1and Plps2containing unsaturated-CH,-C=C-,-CH3,-C-O-C,-OH,-NH2, were both β-glycosidic linkage of the pyran ring polysaccharides; Plps3containing unsaturated-CH,-C=C-,-C-O-C,=CH2,-OH,-CH3,-C=O, was a-glycosidic and β-glycosidic linkage of the pyran ring polysaccharide.The three-component polysaccharides were used to the study of physiological activity, mainly for anti-tumor experiments and immunological experiments on the basis of the above studies. We could see the three kinds of polysaccharides have certain results, but the effects were not significant, through the determination of reducing power and hydroxyl radical scavenging abilities of experiments. In anti-tumor experiment, it showed that Plps-1could inhibit S180cell very well in vitro, inhibition rate up to93.6%. At the same time, Annexin V-FITC/PI double staining flow cytometry identification of cell apoptosis and cell cycle experiment, the experimental results proved that Plps-1could induce apoptosis of S180cells further, and could inhibit the cells arrest at G1phase. In measuring the content of the cytokine TNF-a experiment, we could clearly know, three kinds of polysaccharides from Phellinus linteus slightly inhibited TNF-α generation,it showed that polysaccharides has no toxic and side effects in antitumor. We could find the three Phellinus linteus polysaccharides could significantly enhance its vitality, dose-response relationship in the spleen cells proliferation assays, in which Plps-1could be increased4180.538%at the dose of200μg/mL; Plps-2could increase6843.064%too; Plps-3could increase542.647%too. In the thymus cells proliferation experiments, we knew two kinds of Phellinus linteus polysaccharides which could significantly enhance their vitalities, Plps-1could improve the vitalities9317.217%at the dose of200μg/mL, but improve13232.7%at the dose of100μg/mL; Plps-2could improve the vitalities11215.8%at the dose of200μg/mL; Plps-3could improve the vitalities24.490%at the dose of200μg/mL, were not significant. In macrophages phagocytosis of neutral red experiments, only Plps-1could significantly improve the phagocytosis abilities, Plps-1could increase54.675%at the dose of200μg/mL. In macrophages to secrete NO experiments, three kinds of Phellinus linteus polysaccharides could significantly improve the secretion of macrophages, Plps-1could increase173.590%at the dose of200μg/mL; Plps-2could increase167.436%too; Plps-3could increase108.9744%too. |
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