竹节参皂苷免疫佐剂作用的实验分析-experimental analysis of the effect of panax japonicus saponin as immune adjuvant.docxVIP

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竹节参皂苷免疫佐剂作用的实验分析-experimental analysis of the effect of panax japonicus saponin as immune adjuvant.docx

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竹节参皂苷免疫佐剂作用的实验分析-experimental analysis of the effect of panax japonicus saponin as immune adjuvant

三峡大学硕士学位论文三峡大学硕士学位论文 PAGE IV PAGE IV 万方数据 Abstract Objective: High performance liquid chromatography (HPLC) was determined to screen the effective parts of the immune chikusetsu saponin which has a adjuvant effect, and we studied and discussed this effective part which used HBsAg antigen and adjuvant possible mechanism. Then chikusetsu saponin can as a research and development foundation immune adjuvant. Method: 1 To separate and analysis the preliminary bamboo ginseng saponins by D101 macroporous resin and HPLC; 2 In vitro, we aseptically isolated from mouse spleen lymphocytes, training, administration, and then by MTT assay, to invest the best parts of the immune adjuvant activity of the different concentrations of ethanol eluted bamboo ginseng saponin; 3 Six weeks female Balb / c mice were 54, randomly divided into six groups 9: normal group (Z), antigen group (HBsAg , 5 μg / only), antigen + 60% ethanol bamboo ginseng saponin low-dose group (HBsAg + 60% low, 300 μg / only), antigen + 60% ethanol in the high dose group chikusetsu saponin (HBsAg + 60% high, 600 μg / only), antigen + ginsenosides rg1 low-dose group (HBsAg + rg1 low, 50 μg / only), antigen + ginsenosides rg1 high dose group (HBsAg + rg1 high, 100 μg / only); respectively in the 0, 7 and 21 days, the mice were vaccined, and each group were given saline, HBsAg, HBsAg + 60% low, HBsAg + 60% high, HBsAg + rg1 low and HBsAg + rg1 high. After the end of the immunization of 7 days, the mice were sacrificed by cervical dislocation; (1) By the CCK-8 assay, to invest antigen HBsAg and ConA-induced proliferation of mouse lymphocytes situation; (2) By the indirect ELISA, to invest serum mouse IgG, IgG1 and IgG2b antibody titers; (3) By the RT-PCR, to detect cytokines IL-2, IFN-γ, IL-4 and IL-10 expression levels; (4) By flow cytometry cell, to detect the expression of surface protein TLR4 and TLR9. Results: 1 Initial realization of the separation chikusetsu saponin substances in each segment and ide