JAZF1过表达对糖异生及胰岛素信号通路的影响  

Effect of JAZF1 over-expression on gluconeogenesis and insulin signal pathway

作　　者：王汉生[1] 李伶[1] 杨刚毅[1] 罗小河[1] 李敏燕[1] 代继桓 任艳[1] 

机构地区：[1]重庆医科大学检验医学院临床生化教研室,教育部临床检验诊断学重点实验室,重庆400010

出　　处：《重庆医科大学学报》2015年第7期965-969,共5页Journal of Chongqing Medical University

基　　金：教育部博士点基金项目资助(编号:20105503110002,20125503110003);重庆市自然科学基金重点项目资助(编号:cstc2012 jj B10022)

摘　　要：目的:在课题组前期细胞研究结果的基础上构建锌指并列基因1(juxtaposed with another zinc finger gene 1,JAZF1)转基因动物模型并探讨JAZF1基因过表达对糖异生及胰岛素信号通路的影响。方法:将构建和制备的JAZF1基因片段,经显微注射到受精卵中,将受精卵移植入受体小鼠,产出后经鉴定和繁育得到较纯的JAZF1转基因小鼠。将8周龄转基因小鼠(Tg-JAZF1)和野生(wild type,WT)小鼠随机分为4组喂养:即普食(standard diet,SD)-WT组(n=6)、SD-Tg组(n=6)、高脂(high fat diet,HFD)-WT组(n=6)、HFD-Tg组(n=6)。各组小鼠喂养12周之后测定肝脏组织糖异生的关键基因葡萄糖-6-磷酸酶(glucose-6-phosphatase,G-6pase)和磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxykinase,PEPCK)的m RNA和蛋白表达的变化。Western blot测定肝脏、肌肉、脂肪组织胰岛素信号通路蛋白磷酸化水平变化。结果:SD-Tg小鼠的m RNA和蛋白在肝脏、肌肉、脂肪组织中表达较SD-WT小鼠均明显升高,差异具有统计学意义(P<0.05);与WT小鼠比较,Tg-JAZF1小鼠肝脏PEPCK、G-6-Pase在SD喂养和HFD喂养条件下m RNA水平和蛋白表达水平均明显降低,差异具有统计学意义(P<0.05);肝脏组织胰岛素信号通路蛋白磷酸化水平均增加,差异具有统计学意义(P<0.05)。结论:JAZF1转基因动物模型构建成功,JAZF1过表达能抑制肝脏糖异生关键基因表达,增加肝脏组织的胰岛素敏感性。Objective：To establish juxtaposed with another zinc finger gene 1（JAZF1）-transgenic mice model on the basic of cell research in vitro and to explore the effect of JAZF1 over-expression on gluconeogenesis and insulin signal pathway. Methods：The prepared JAZF1 gene fragment was microinjected into one-cell embryos of mice,then the JAZF1-transgenic mice were generated and identified by standard procedures. Eight-week-old Tg-JAZF1（Tg） mice and wild type（WT） mice were randomly divided into 4 groups：standard diet（SD）-WT group（n=6）,SD-Tg group（n=6）,high fat diet（HFD）-WT group（n=6） and HFD-Tg group（n=6）. Phosphoenolpyruvate carboxykinase and glucose-6-phosphatase in liver were detected by PCR and Western blot after 12 weeks. The phosphorylation of insulin signal protein was measured by Western blot. Results：JAZF1 m RNA and protein expressions in liver,muscle and adipose tissue were elevated significantly in SD-Tg mice than in SD-WT mice（P0.05）.The expressions of gluconeogenesis gene in Tg mice were dramatically decreased compared with those of WT mice under the condition of SD or HFD（P0.05）. The phosphorylation of insulin signal protein in liver,muscle and adipose tissue were increased in Tg mice than in WT mice（P0.05）. Conclusion：Transgenic mice model has been generated successfully. JAZF1 overexpression inhibit the expression of gluconeogenesis gene and also improve the insulin sensitivity of liver,muscle and adipose tissue.

关 键 词：锌指并列基因1 转基因鼠 磷酸烯醇式丙酮酸羧化酶 葡萄糖-6-磷酸酶 胰岛素信号 

分 类 号：R587.1[医药卫生—内分泌]