Ginsenoside Rg1 And Its Metabolite Rh1 On The Function Of Dendritic Cells And Its Mechanism Research

Dendritic cell (DC) plays important roles not only in taking antigen but also in activating T cell. It is believed that DC is a pivotal Antigen Presenting Cell (APC) in the immune system. In this dissertation, we compared the effects of Ginsenoside Rg1 and its metabolite, Ginsenoside Rh1, on dendritic cell, then elucidated the immuno-regulating mechanism of Ginsenoside Rg1. 1. Separating pregnior DC from PBMC, then inducing these cells with IL-4, GM-CSF and TNFa. 7 days later, identifying mature DC by the combination of microscope, electro-microscope and immunohistochemistry staining with S-100 protein. The results showed that the mature rate of DC is 95-99%。2. The adhesion rate between pregnior DC and HUVEC was evaluated after treatment with Rg1 and Rh1 separately. (1) After 2h treatment with Rg1 and Rh1, HUVEC was cultured with pregnior DC. The results after 24 hours co- cultivation showed that Rg1 100(g/ml and all dose of Rh1 could enhance the adhesion rate between pregnior DC and HUVEC with and without stimulation of TNFa, moreover the effect of Rh1 1,10(g/ml is more significant than the same concentration of Rg1 (P<0.001). the results after 48h co-cultivation showed that all dose of Rh1 could promote the adhesion rate in the lack of stimulator and the effect is more significant than the same concentration of Rg1 (P<0.001); with TNFαas stimulator, Rh1 10,100(g/ml could enhance significantly the adhesion rate(P<0.05-0.01)and the effect is more significant than the same concentration of Rg1 (P<0.001).(2) After 2h treatment with Rg1 and Rh1, pregnior DC was cultured with HUVEC. The results After 24h co-cultivation showed that both Rg1 and Rh1 could enhance significantly the adhesion rate between pregnior DC and HUVEC (P<0.001) with and without the stimulation of TNFα; The results after 48h co-cultivation showed that only Rg1 10(g/ml and Rh1 100(g/ml could improve the adhesion rate (P<0.05-0.01). With TNFαas stimulator, all dose of Rh1 could improve the adhesion rate (P<0.01-0.001), however Rg1 10,100(g/ml showed obvious inhibitory effect on adhesion (P<0.001).3. The effect Rg1 and Rh1 on the proliferation of T cell stimulated by DC was detected using MTT method. The result demonstrated that Rh1 10(g/ml play inhibitory and stimulating effect on the proliferation of T cell when the ratio between T cell and DC was 10:1 and 25:1 separately(P<0.05).Both Rg1 and Rh1 have no effect on the proliferation of T cell when the ratio of TC and DC is 50:1.4. The cytotoxicity activity of DC-LPAK was detected using neutral red phagocytosis assay. The results showed that both Rg1 and Rh1 could enhance the cytotoxicity activity of DC-LPAK on papilla tumor cell line; When the E:T was 5:1, all dose of Rg1 could enhance the cytotoxicity of DC-LPAK on L929 cell line(P<0.01；0.05),. However only 10μg/ml Rh1 can improve the cytotoxicity of DC-LPAK (P<0.05).5. The immunohistochemistry results showed that both Rg1 and Rh1 could stimulate the expression of surface molecules of DC, such as HLA-DR,CD25, HLA-DR, CD44, CD54 and CD11c. However they have no significant effect on the expression of E-seletctin.6. The effect of Rg1 and Rh1 on the expression and transcription lever of IL-12 P40 was detected using ELISA and RT-PCR.. The result showed that all dose of Rg1 and Rh1 could enhance the expression of IL-12 P40. Rg1 100, 1(g/ml and Rh1 100(g/ml could improve the transcription lever the IL-12 P40 mRNA and the effect of Rg1 100μg/ml is more significant.The results demonstrated that both Rg1 and its metabolite Rh1 could enhance the migration of precursor DC, they can improve the proliferation of T cell and enhance the cytotoxity activity of LPAK by improving the expression of adhesion and co-stimulating moleculars on the surface of mature DC and the transcription and translation of IL-12 . The result showed that the immune effects of Rg1 was proformed by Rg1 and Rh1 which were absorbed into blood.