脂联素对小鼠免疫性肝损伤保护作用的研究
发布时间：2019-01-06 14:42
【摘要】：脂联素是近来发现的一种脂肪组织特异性产生的细胞因子,它除了调节糖脂代谢、改善胰岛素抵抗外,还能通过抑制肿瘤坏死因子α(TNF-α)的产生与作用来调节机体的炎症反应。病毒性肝炎的主要致病机理就是活化的T细胞产生以TNF-α为主的细胞因子介导对肝细胞的免疫性损伤。因此,本文采用Con A诱导性肝损伤小鼠模型进行脂联素干预实验,以探讨脂联素是否对肝脏的免疫性损伤具有保护作用。研究的主要内容包括:①从脂肪组织克隆小鼠脂联素基因,并在大肠杆菌内表达;②构建小鼠脂联素真核表达载体pAA-neo-mAd,并在COS-7细胞中验证其表达;③将表达载体pAA-neo-mA与pCI-neo-mAd导入小鼠体内,验证它们在肝内的表达及表达效率;④在将载体pAA-neo-mAd导入小鼠体内,并获得重组脂联素在肝脏内的高效稳定表达的基础上,进行Con A致小鼠免疫性肝损伤的脂联素干预实验,通过肝脏组织学检查、血清ALT、TNF-α浓度测定等比较表达载体组与空载体组(阴性对照)肝损伤程度的不同,并分析脂联素、TNF-α及ALT之间的相关性。 研究结果发现,表达载体组小鼠的肝脏损伤明显轻于空载体组(两组ALT比较,P0.0001),其血清TNF-α浓度也低于空载体组(P0.0001)。初步表明脂联素对Con A导致的肝脏免疫性损伤具有保护作用,其作用机制可能是通过抑制TNF-α的产生。 本研究将流体动力法(Hydrodynamics-based procedure)应用于小鼠脂联素的体内表达,成功地将脂联素真核表达载体导入了小鼠肝脏,获得了重组脂联素在肝内的高效稳定表达。并通过脂联素对Con A致肝脏免疫性损伤的干预实验初步表明,脂联素对Con A导致的T细胞介导的肝脏免疫性损伤具有保护作用。这一发现为脂联素在病毒性肝炎临床防治方面的研究提供了重要的理论依据。
[Abstract]:Adiponectin is a recently discovered cytokine specifically produced by adipose tissue. In addition to regulating glucose and lipid metabolism, adiponectin improves insulin resistance. It can also regulate the inflammatory response by inhibiting the production and action of tumor necrosis factor 伪 (TNF- 伪). The main pathogenetic mechanism of viral hepatitis is the immune damage induced by activated T cells by cytokines mediated by TNF- 伪. In order to investigate whether adiponectin has protective effect on immune injury of liver, adiponectin was used to induce liver injury in Con A mice. The main contents of this study were as follows: (1) the adiponectin gene was cloned from adipose tissue and expressed in E. coli; (2) the eukaryotic expression vector of mouse adiponectin (pAA-neo-mAd,) was constructed and its expression was verified in COS-7 cells. (3) the expression vectors pAA-neo-mA and pCI-neo-mAd were introduced into mice to verify their expression and expression efficiency in liver. 4 on the basis of introducing the vector pAA-neo-mAd into mice and obtaining the high and stable expression of recombinant adiponectin in the liver, the adiponectin intervention experiment of immune liver injury induced by Con A in mice was carried out. The liver histological examination and serum ALT, were carried out. The degree of liver injury was compared between the expression vector group and the empty vector group (negative control group), and the correlation among adiponectin, TNF- 伪 and ALT was analyzed. The results showed that the liver injury in the expression vector group was significantly lighter than that in the empty vector group (P 0.0001), and the serum TNF- 伪 concentration was lower in the expression vector group than in the empty vector group (P 0.0001). It is suggested that adiponectin has protective effect on hepatic immune injury induced by Con A, and its mechanism may be by inhibiting the production of TNF- 伪. In this study, the hydrodynamic method (Hydrodynamics-based procedure) was applied to the expression of adiponectin in mice. The eukaryotic expression vector of adiponectin was successfully introduced into mouse liver, and the high and stable expression of recombinant adiponectin in the liver was obtained. The intervention of adiponectin on Con A induced liver immune injury showed that adiponectin had protective effect on T cell mediated hepatic immune injury induced by Con A. This discovery provides an important theoretical basis for the study of adiponectin in clinical prevention and treatment of viral hepatitis.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392
本文编号：2402929
[Abstract]:Adiponectin is a recently discovered cytokine specifically produced by adipose tissue. In addition to regulating glucose and lipid metabolism, adiponectin improves insulin resistance. It can also regulate the inflammatory response by inhibiting the production and action of tumor necrosis factor 伪 (TNF- 伪). The main pathogenetic mechanism of viral hepatitis is the immune damage induced by activated T cells by cytokines mediated by TNF- 伪. In order to investigate whether adiponectin has protective effect on immune injury of liver, adiponectin was used to induce liver injury in Con A mice. The main contents of this study were as follows: (1) the adiponectin gene was cloned from adipose tissue and expressed in E. coli; (2) the eukaryotic expression vector of mouse adiponectin (pAA-neo-mAd,) was constructed and its expression was verified in COS-7 cells. (3) the expression vectors pAA-neo-mA and pCI-neo-mAd were introduced into mice to verify their expression and expression efficiency in liver. 4 on the basis of introducing the vector pAA-neo-mAd into mice and obtaining the high and stable expression of recombinant adiponectin in the liver, the adiponectin intervention experiment of immune liver injury induced by Con A in mice was carried out. The liver histological examination and serum ALT, were carried out. The degree of liver injury was compared between the expression vector group and the empty vector group (negative control group), and the correlation among adiponectin, TNF- 伪 and ALT was analyzed. The results showed that the liver injury in the expression vector group was significantly lighter than that in the empty vector group (P 0.0001), and the serum TNF- 伪 concentration was lower in the expression vector group than in the empty vector group (P 0.0001). It is suggested that adiponectin has protective effect on hepatic immune injury induced by Con A, and its mechanism may be by inhibiting the production of TNF- 伪. In this study, the hydrodynamic method (Hydrodynamics-based procedure) was applied to the expression of adiponectin in mice. The eukaryotic expression vector of adiponectin was successfully introduced into mouse liver, and the high and stable expression of recombinant adiponectin in the liver was obtained. The intervention of adiponectin on Con A induced liver immune injury showed that adiponectin had protective effect on T cell mediated hepatic immune injury induced by Con A. This discovery provides an important theoretical basis for the study of adiponectin in clinical prevention and treatment of viral hepatitis.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392
【参考文献】
相关期刊论文 前1条
1 ;Insulin expression in livers of diabetic mice mediated by hydrodynamics-based administration[J];World Journal of Gastroenterology;2004年04期
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