siRNA沉默HIF-1α对大鼠心肌细胞CT-1表达的影响  

Effect of silencing hypoxia inducible factor 1α by siRNA on the expression of cardiotrophin-1 in rat myocardium cell

作　　者：王晗[1] 符史干[1] 董战玲[1] 许闽广[1] 王杨[1] 陈国斌[1] 高凌峰[1] 

机构地区：[1]海南医学院生理学教研室,海南海口571199

出　　处：《基础医学与临床》2013年第1期88-92,共5页Basic and Clinical Medicine

基　　金：海南省自然科学基金(309035);海南省教育厅高校科研基金(Hjkj2009-55)

摘　　要：目的研究低氧诱导因子-1α(HIF-1α)在心肌细胞适应低氧环境中作用机制。方法合成针对HIF-1α的siRNA片段,转染大鼠心肌细胞系H9C2。Real-time PCR检测HIF-1α和心肌营养素(CT-1)mRNA水平,Westernblot检测HIF-1α和CT-1蛋白水平。结果低氧环境下,转染针对HIF-1αsiRNA片段后HIF-1αmRNA水平、HIF-1α蛋白质水平、CT-1 mRNA水平和蛋白水平均明显减低(P<0.05)。不进行转染时,CT-1 mRNA水平、HIF-1α蛋白和CT-1蛋白水平在低氧下比常氧下明显增高(P<0.05)。CT-1蛋白由对照组的0.34±0.05增至0.55±0.05(P<0.05)。结论低氧情况下CT-1基因的调控作用有可能是通过低氧激活HIF-1α而诱导产生的,HIF-1α在心肌细胞适应低氧环境中具有重要作用。Objective To study the regulation mechanism of hypoxia-inducible factor-1α（HIF-1α）in rat myocardium cell under hypoxia environment.Methods Small interference RNA（siRNA） targeting at HIF-1α was constructed and transfected into rat myocardium cell lines（H9C2）.The HIF-1α and cardiotrophin-1（CT-1）gene expression at mRNA levels were detected by RT-PCR.The HIF-1α and CT-1 gene expression at protein levels were detected by western blot.Results Expression of the mRNA and protein level of HIF-1α and CT-1 were down regulated after silencing of HIF-1α under hypoxia environment（P0.05）.Expression of the protein level of HIF-1α and the mRNA and protein level of CT-1 increased after exposure of untransfected H9C2 cells to hypoxia（P0.05）.Expression of the protein level of CT-1 increased from 0.34±0.05 to 0.55±0.05（P0.05）.Conclusions The regulation of CT-1 gene under hypoxia could be activated by HIF-1α that plays an important role in myocardial adaptation to hypoxia.

关 键 词：siRNA片段 低氧诱导因子-1Α 心肌营养素-1 心肌低氧 

分 类 号：R363[医药卫生—病理学]