【摘要】： 甘氨膽酸是膽汁酸的主要成分之一,屬于結(jié)合型膽汁酸,在肝臟中由膽酸和甘氨酸結(jié)合而成。多種動(dòng)物膽汁中均含有一定比例的甘氨膽酸。為了探索從牛膽汁中分離提取甘氨膽酸的工藝路線,并了解甘氨膽酸對(duì)機(jī)體免疫功能的影響。采用鹽析、萃取、柱層析等生物化學(xué)方法,從牛膽汁中提取出了有效成分甘氨膽酸,并采用薄層層析法及薄層掃描法進(jìn)行定量、定性檢測(cè),測(cè)得甘氨膽酸的純度可達(dá)98%以上,牛膽汁甘氨膽酸的回收率為64%。采用sc環(huán)磷酰胺(CTX)及環(huán)孢菌素A(Cyclosporin A CsA)制造免疫抑制模型;采用ELISA及比色法觀察GCA對(duì)體液免疫的影響;碳廓清法測(cè)定了GCA對(duì)單核巨噬細(xì)胞的吞噬功能的影響;采用遲發(fā)型皮膚變態(tài)反應(yīng)法觀察GCA對(duì)二硝基氯苯(DNCB)丙酮溶液所致小鼠遲發(fā)型皮膚過敏反應(yīng)的影響,并測(cè)定了血清中IL-2、TNF-α的含量;采用流式細(xì)胞術(shù)檢測(cè)了GCA對(duì)小鼠外周血T細(xì)胞亞群CD4+、CD8+比例的影響。結(jié)果顯示GCA可以顯著增強(qiáng)正常及抑制小鼠的IgM、IgG的含量;顯著提高巨噬細(xì)胞的吞噬功能;顯著增加外周血中T淋巴細(xì)胞CD4+比例,使CD4+/CD8+比值提高;顯著抑制遲發(fā)型皮膚變態(tài)反應(yīng)的發(fā)生,并顯著增加IL-6和TNF-α的含量。表明,GCA對(duì)機(jī)體免疫功能具有調(diào)節(jié)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- 偽 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- 偽. The results showed that GCA could regulate the immune function of the body.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

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