Studies On Extraction And Purification Techniques And Immunological Effects Of Glycocholic Acid

Glycocholic acid (GCA) is a kind of glycol-conjugated bile acid which exists in many animals'bile. It is one of main component of bile acid and was synthesized from cholalic acid and glycine in liver. The aim of the study is to explore extraction and purification techniques and immunological effects of GCA.GCA was extracted and purified from bovine bile by using the biochemistry techniques such as salting-out,extracted,dehydration,silica gel separation. The qualitative and quantitative analysis of GCA were performed by using thin-layer chromatography(TLC).The purity of GCA was over 98%,and reclaim rate was 64%.Suppressive model of mice were made by CTX and CsA. The effect of GCA on humoral immunity were tested by using ELISA and turbidimetric method. The phagocytosis of mononuclear phagocyte system was determined by using carbon particle clearance test. The influence of GCA on levels of delayed-type hypersensivity(DTH) induced by 7% DNCB was tested by using delayed type hypersensitivity and cellular factors IL-2,TNF-αwere observed by using ELISA. In the mean while, the index of immune organs were weighed and calculated. The expression rate of CD4+,CD8+ of T-Cell subgroups were analyzed by using Flow cytometry. The results showed that GCA significantly increased the content of IgM,IgG in normal and suppressive mice, markedly improve the phagocytosis functions of mononuclear phagocyte system in mice. GCA restrained level of DTH remarkably, it also enhanced level of serum IL-2 and TNF-αsignificantly. The percentage of CD4+ and CD4+/CD8+ value in peripheral blood were significantly enhanced. These results suggest that GCA can regulate immune function in mice.