Study On Effect Of Proliferation And Apoptotic In Immune Organs And RAW264.7Cell Lines By TCDCA In Mice

The researches about bile acid and receptor have become a hotspot at domestic and foreign at present. In recent study a variety pharmacological effects of TCDCA through receptors of TGR5and FXRa were found, such as regulating endocrine, immune function and ability of metabolic and so on. TCDCA is an important active ingredient of bile acid, our previous studies have shown that TCDCA have a effect of regulating immune function in mice. In order to further reveal mechanism of immunoregulation of TCDCA in mice, we carried out the research of proliferation and apoptotic in immune organs of mice and RAW264.7cells by TCDCA. The main research contents and results are as follows:In vivo experiments, the mices were divided randomly into four groups:control group, normal and TCDCA (0.1g/kg) group, model group, model and TCDCA(0.1g/kg) group. Immune organs pathological changes were observed through paraffin section and hematoxylin and eosin stain (HE), immune cells proliferation effect were observed through EDU, immune cells apoptosis were observed through TUNEL. HE results showed, compared with the model group, the pathological changes of TCDCA group were more obviously. Compared with the control group, the cells proliferation index of normal and TCDCA group were significantly improved. Compared with the model group, the cells proliferation index of model and TCDCA group were significantly improved, the proliferation of immune cells was obviously improved by TCDCA. TUNEL results showed, compared with the model group, cells apoptosis index of model and TCDCA group were significantly decreased. TCDCA can obviously resist the apoptosis cased by DEX.In vitro experiments, RAW264.7cell proliferation effects were observed through CCK-8reagent kit. The results indicated that TCDCA could obviously promote cells proliferation, and cells proliferation were more obviously together with LPS. The expressions of cyclins mRNA were measured by realtime fluores-cence quantitative PCR, the results indicated that the mRNA expressions of CyclinDl, CyclinEl, CyclinAl and CDK2were different degree increased by different TCDCA dose. The diphenylamine experiment was used for examining the percentage of apoptosis in RAW264.7, the results indicated that TCDCA could decrease the percentage of DNA fragmentation compared with the model group, TCDCA could also prevent cells apoptosis that were induced by DEX, but this depended on different doses of TCDCA. The mRNA expressions of IAPs were measured by using RTFQ-PCR, the results indicated that the mRNA expressions of CIAP-1, CIAP-2and XIAP were different degree increased by different TCDCA dose.These findings suggest that TCDCA can promote cells proliferation and resist the cells apoptosis of immune organs and RAW264.7cells in mice. At the same time, the mRNA expressions of Cyclins and IAPs are different degree increased by different TCDCA dose.