甘氨胆酸的提取纯化及其对小鼠免疫作用的研究

发布时间：2019-05-23 09:19

【摘要】： 甘氨胆酸是胆汁酸的主要成分之一,属于结合型胆汁酸,在肝脏中由胆酸和甘氨酸结合而成。多种动物胆汁中均含有一定比例的甘氨胆酸。为了探索从牛胆汁中分离提取甘氨胆酸的工艺路线,并了解甘氨胆酸对机体免疫功能的影响。采用盐析、萃取、柱层析等生物化学方法,从牛胆汁中提取出了有效成分甘氨胆酸,并采用薄层层析法及薄层扫描法进行定量、定性检测,测得甘氨胆酸的纯度可达98%以上,牛胆汁甘氨胆酸的回收率为64%。采用sc环磷酰胺(CTX)及环孢菌素A(Cyclosporin A CsA)制造免疫抑制模型;采用ELISA及比色法观察GCA对体液免疫的影响;碳廓清法测定了GCA对单核巨噬细胞的吞噬功能的影响;采用迟发型皮肤变态反应法观察GCA对二硝基氯苯(DNCB)丙酮溶液所致小鼠迟发型皮肤过敏反应的影响,并测定了血清中IL-2、TNF-α的含量;采用流式细胞术检测了GCA对小鼠外周血T细胞亚群CD4+、CD8+比例的影响。结果显示GCA可以显著增强正常及抑制小鼠的IgM、IgG的含量;显著提高巨噬细胞的吞噬功能;显著增加外周血中T淋巴细胞CD4+比例,使CD4+/CD8+比值提高;显著抑制迟发型皮肤变态反应的发生,并显著增加IL-6和TNF-α的含量。表明,GCA对机体免疫功能具有调节作用。
[Abstract]:Glycine cholic acid is one of the main components of bile acid, which belongs to bound bile acid and is formed by the combination of cholic acid and glycine in the liver. Many kinds of animal bile contain a certain proportion of glycine cholic acid. In order to explore the separation and extraction of glycine cholic acid from bovine bile and to understand the effect of glycine cholic acid on immune function. Glycine cholic acid was extracted from bovine bile by salting out, extraction and column chromatography. The purity of glycine cholic acid was over 98% by thin layer chromatography and thin layer scanning. The recovery rate of glycine cholic acid in bovine bile was 64%. The immunosuppressive model was established by sc cyclophosphamide (CTX) and cyclosporine A (Cyclosporin A CsA), the effect of GCA on humoral immunity was observed by ELISA and colorimetric method, and the effect of GCA on the phagocytosis of monocytes was measured by carbon profile assay. The effect of GCA on delayed skin allergic reaction induced by dinitrochlorobenzene (DNCB) acetone solution in mice was observed by delayed skin allergy method, and the content of IL-2,TNF- 伪 in serum was determined. The effects of GCA on the ratio of CD4 and CD8 in peripheral blood T cell subsets of mice were detected by flow cytometry. The results showed that GCA could significantly enhance the content of IgM,IgG and inhibit the content of IgM,IgG in mice, significantly increase the phagocytic function of macrophages, significantly increase the CD4 ratio of T lymphocytes in peripheral blood, and increase the ratio of CD4 / CD8. It significantly inhibited the occurrence of delayed skin allergy and significantly increased the contents of IL-6 and TNF- 伪. The results showed that GCA could regulate the immune function of the body.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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