利用体外合成mRNA方法研究Betatrophin对糖代谢关键基因表达的影响  

Study on the Molecular Mechanism of Betatrophin Regulated Glucose Metabolism by Synthesis of Betatrophin mRNA in vitro

作　　者：汪兰[1,2] 付红霞[1] 郭兴荣[1] 

机构地区：[1]十堰市太和医院.湖北医药学院附属医院胚胎干细胞湖北省重点实验室 [2]湖北医药学院药学院,湖北十堰442000

出　　处：《湖北医药学院学报》2016年第6期531-535,共5页Journal of Hubei University of Medicine

基　　金：2016年湖北省自然科学基金面上项目(2016CFB408);国家自然科学基金应急管理项目(81641028);湖北省卫生计生委一般项目(WJ2015MB223);2015年湖北医药学院药学院本科生科研训练计划(SRTP)项目

摘　　要：目的:利用体外合成mRNA的方法研究新基因Betatrophin在HepG2细胞中对糖代谢关键基因表达的影响。方法:以合成Betatrophin mRNA的质粒为模板,体外合成Betatrophin mRNA转染入HepG2细胞,利用免疫荧光和Western blot检测Betatrophin mRNA在细胞内Betatrophin蛋白过表达情况,再利用qPCR和Western blot检测Betatrophin过表达对糖代谢关键基因表达影响。结果:成功构建Betatrophin mRNA合成载体,体外合成的Betatrophin mRNA可高效转染到HepG2细胞株中并正确表达Betatrophin蛋白,高表达的Betatrophin可上调糖代谢关键基因Glut2表达。结论:本实验初步证实Betatrophin可能是通过调控Glut2表达来影响糖代谢,为进一步研究Betatrophin对调控糖代谢的分子作用机制打下基础。Objective To study the molecular mechanism of betatrophin regulating glucose metabolism in HepG2 cell by synthesis of betatrophin mRNA in vitro.Methods Using the synthetic betatrophin mRNA plasmid as template, transfected synthetic betatrophin mRNA into HepG2 cell and detected the expression of betatrephin protein in cells by using immunofluorescence and Western blot, then analyzed the effect of betatmphin over-expression on glucose metabolism related genes expression by qPCR and Western blot. Results Betatrophin mRNA plasmid was successfully constructed, the synthetic betatrophin mRNA in vitro could be efficiently transfected into HepG2 cells and correctly expressed betatrophin protein, the high expression of betatrophin could up regulate the key glucose metabolism gene Glut2 expression. Conclusion This experiment confirmed that betatrophin may affect theglucose metabolism through regulating the expression of Glut2, which lays a foundation for further study on the mechanism of molecular of betatrophin on the regulation of glucese metabolism.

关 键 词：Betatrophin HEPG2细胞株 糖代谢 GLUT2 

分 类 号：R587.1[医药卫生—内分泌]