基础医学与临床 ›› 2013, Vol. 33 ›› Issue (1): 88-92.

• 研究论文 • 上一篇    下一篇

siRNA沉默HIF-1α对大鼠心肌细胞CT-1表达的影响

王晗1,符史干2,董战玲2,许闽广2,王杨2,陈国斌2,高凌峰1   

  1. 1. 海南医学院生理学教研室
    2. 海南医学院
  • 收稿日期:2012-03-05 修回日期:2012-05-30 出版日期:2013-01-05 发布日期:2012-12-25
  • 通讯作者: 王晗 E-mail:wanghan2801@sohu.com
  • 基金资助:
    海南省自然科学基金;海南省教育厅高校科研基金

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

  • Received:2012-03-05 Revised:2012-05-30 Online:2013-01-05 Published:2012-12-25

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,Western blot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-1mRNA水平和蛋白水平均明显减低(P<0.05)。不进行转染时,CT-1mRNA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。

关键词: siRNA片段, 低氧诱导因子-1α, 心肌营养素-1, 心肌低氧

Abstract: 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) targeing 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 real-time 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(P<0.05). 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(P<0.05). Expression of the protein level of CT-1 increased from 0.34±0.05 to 0.55±0.05(P<0.05).Conclusion The regulation of CT-1 gene under hypoxia could be activated by HIF-1α that plays an important role in myocardial adaptation to hypoxia.

Key words: small interference RNA, hypoxia-inducible factor-1α, cardiotrophin-1, myocardium under hypoxia environment.

中图分类号: