The Study Of Immunological Activity Of Polysaccharides Extracted From Hericium Erinaceus On Murine Splenic Lymphocytes And Macrophage RAW264.7 Cells In Vitro

Objectives:1.To investigate the effects of HEP on murine splenic lymphocyte proliferation,cell cycle,the secretion and mRNA expression of Thl cytokines(IL-2,IFN-?,TNF-?),Th2 cytokines(IL-4,IL-6),NO in vitro.2.To investigate the effects of HEP on RAW264.7 cells proliferation,cell cycle,cell apoptosis,phagocytic function,the secretion of TNF-??IL-1??IL-6?IL-10?NO and the xpression of TNF-??IL-1??IL-6?IL-10?iNOS mRNA and the expression of protein NF-?B.Methods:1.The cell survival rates were assessed by MTT;The effects of HEP on the cell cycle were detected by flow cytometry;The secretion of Thl cytokines(IL-2,IFN-y,TNF-a),Th2 cytokines(IL-4,IL-6),NO in the supernate were determined by ELISA;the gene expression of Thl cytokines(IL-2,IFN-?,TNF-?),Th2 cytokines(IL-4,IL-6),T-bet GATA-3,iNOS were determined by RT-PCR.2.The cell survival rates were assessed by MTT;The effects of HEP on the cell cycle and cell apoptosis were detected by flow cytometry;The secretion of TNF-a,IL-1p,IL-6,IL-10 and NO in the supernate were determined by ELISA;the gene expression of TNF-a,IL-1?,IL-6,IL-10 and iNOS mRNA were determined by RT-PCR.Results:1.HEP could effectively increase the mouse spleen lymphocyte and T,B lymphocyte proliferation(P<0.05).The stimulation of HEP could induce the mouse spleen lymphocyte and T,B lymphocyte from G0/G1 period to S period;HEP could elevate the levels of Thl cytokines(IL-2,IFN-y,TNF-a),Th2 cytokines(IL-4,IL-6)and NO on the mouse spleen lymphocyte and T lymphocyte;HEP could elevate the gene expression of Thl cytokines(IL-2,IFN-y,TNF-a),Th2 cytokines(IL-4,IL-6),transcription factors(T-bet,GATA-3)and iNOS on the mouse spleen lymphocyte and T lymphocyte.2.HEP could effectively increase RAW264.7 cells proliferation(P<0.05);HEP could induce RAW264.7 cells from G0/G1 period to S period;HEP could inhibit RAW264.7 cell apoptosis;HEP could elevate the levels of TNF-?,IL-1?,IL-6,IL-10 and NO;HEP could elevate the gene expression of TNF-a,IL-1?,IL-6,IL-10 and iNOS.Conclusions:1.HEP could enhance spleen lymphocyte cells proliferation through promoting the synthesis of DNA and could stimulate the cytokine production through elevating the gene expression that HEP could strong immunoenhancement activity on spleen lymphocyte.2.HEP could enhance RAW264.7 cells proliferation through promoting the synthesis of DNA and inhibiting cell apoptosis,also could stimulate the cytokine production through elevating the gene expression,So that HEP could strong immunoenhancement activity on RAW264.7 cells.