大鼠小肠分泌胰岛素基因水平研究  

Research on gene level of small intestine secreting insulin in rats

作　　者：甘文学 张瑞[1] 张欣[1] 汪蓓蕾[1] 郭刚[1] 

机构地区：[1]天津医科大学代谢病医院内分泌研究所,卫生部激素与发育重点实验室,天津300070

出　　处：《天津医科大学学报》2014年第2期98-101,共4页Journal of Tianjin Medical University

摘　　要：目的:明确小肠内胰岛素的存在情况及其表达水平。方法:8周龄雄性Wister大鼠,禁食24 h,灌胃50%葡萄糖溶液、牛血清白蛋白溶液、油脂悬液各2 mL,分别灌胃后在第0、5、10、15、20、25 min时脱臼处死,按不同时间段分为6组,取其胰腺、十二指肠。提取各组织总RNA,反转录成cDNA,普通PCR检测胰岛素mRNA的存在,实时荧光定量PCR检测不同时刻胰腺和十二指肠胰岛素mRNA的相对表达量。结果:反转录普通PCR在大鼠十二指肠中扩增出胰岛素基因片段(经测序证实与胰岛素基因序列完全一致);十二指肠与胰腺胰岛素mRNA的相对表达量有显著差异(P<0.05),远远低于胰腺组织。灌胃后胰腺和十二指肠分泌胰岛素的表达水平呈动态表化,且十二指肠分泌胰岛素的表达高峰晚于胰腺。结论:该实验证实了除胰岛β细胞外十二指肠组织具有分泌胰岛素的功能,灌胃后十二指肠胰岛素mRNA的相对表达量呈动态表化,从而推测糖尿病的一种可能发病机制。Objective： To identify the existence and the expression level of small intestine insulin mRNA. Methods： The Wister rats by irrigation with 50% glucose solution, bovine serum albumin solution and oil suspension were executed at 0,5,10,15,20,25 rain, according to which the rats were divided into six groups before the sample of pancreas and duodenum were obtained. Total RNA were extracted from tissues. Reverse transcriptase PCR detected the existence of insulin mRNA in each tissue and real-time fluorescence PCR detected insulin mRNA expression in each part of duodenum and pancreas. Results： Reverse transcriptase PCR amplified gene sequencing in the small intestine insulin （completely consistent with insulin genetic sequences）. Based on detection by real-time fluorescent quantitative PCR, the insulin mRNA relative expression in duodenum was significantly lower than that in pancreas （P〈0.05）. The insulin expression levels in duodenum and pancreas were dynamic, and the insulin expression peak of the duodenum was later than that of pancreas. Conclusion： The research shows that the small intestine tissues secrete insulin except for pancreatic islet β cell The insulin expression levels in duodenum and pancreas are dynamic, and thus can be viewed as a possible pathogenesis for diabetes.

关 键 词：小肠 胰岛素 肠源胰岛素 大鼠 

分 类 号：R587.1[医药卫生—内分泌]