| In this study,fruiting body of Ganoderma tsugae was extracted by hot water,and was precipitated by alcohol.The Ganoderma Tsugae polysaccharide(GT)was obtained,GT was brown powder,the polysaccharide of GT is 56.74%,the protein of GT is 40.8%,and the yield of GT is 2,1%.Further research about separation,purification and identification of properties of GT were operated.GT was separated into two components(GTW and GTN)by DEAE-Sepharose Fast Flow ion exchange chromatography.The content of polysaccharide is determined by phenol-acid method and the content of protein is Folin-phenol assay in the same tube.The results showed,GTW is water-soluble protein-polysaccharide,is creamy white powder.The content of polysaccharide of GTW was 74.84%,and the content of protein of GTW was 20.4%.The yield of GTW was up to 0.5%(relative to the raw medicinal herbs).The datas of HPLC reflected that molecular weight of GTW is 7000-10000Da.GTW has two components.Then GTW was further purified by Sephedex G-100 column.GTWa was obtained and the yield was 0.1%(relative to the raw medicinal herbs).GTWa was single molecular weight of the protein-polysacharides which molecular weight was 7896.8Da by HPLC.Infrared spectroscopy analysis showed that GTW was ?-glycoside proteoglycan.Suger analysis revealed that GTW was composed of mannose(Man),glucose(Glc)and galactose(Gal):Area percent Man:Glc:Gal=56.93:23.30:10.15.The liver protective effect and lowering blood lipid activity of GTW were screened on HepG2 cells,human liver hepatocellular carcinoma cell line.The results showed that different concentrations of GTW could improve the survival rate of HepG2 cells which were damaged by alcohol.GTW could inhibit ALT,AST,MDA expression,improve SOD activity,reduce the number of apoptotic on HepG2 cells.GTW could increase LDLR protein expression of HepG2 cells.The results showed that GTW,proteoglyean of Ganoderma tsugae,has liver protection and lipid-lowering activity. |
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