在模拟低氧环境下心肌细胞HIF-1与CT-1的相互作用  

Interaction between HIF-1 and CT-1 in rat myocardium cell line under hypoxia environment

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：《海南医学院学报》2013年第3期289-292,共4页Journal of Hainan Medical University

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:研究低氧环境下大鼠心肌细胞系H9C2细胞中HIF-1与CT-1的相互作用。方法:氯化钴(CoCl2)模拟低氧环境。采用CCK-8试剂盒检测心肌细胞存活率。化学合成siRNA片段,并通过li-pofectamine 2 000转染大鼠心肌细胞系H9C2。采用Real-time PCR和Western-blot技术分别检测HIF-1α和CT-1的mRNA和蛋白质水平。结果:低氧处理48h后,干扰组(转染针对HIF-1α的siR-NA的H9C2)中CT-1mRNA和蛋白水平较对照组(转染无义干扰片段的H9C2)明显减少(P<0.05)。低氧3d和4d后,加入CT-1组的心肌细胞存活率较对照组高(P<0.05),HIF-1mRNA表达水平与对照组无显著差异(P>0.05),HIF-1蛋白水平增加。结论:在培养心肌细胞中模拟低氧环境,HIF-1表达减少伴随着CT-1表达减少,加入CT-1后,HIF-1蛋白表达也增加,HIF-1与CT-1相互作用。Objective： To investigate the interaction between HIF-1 （hypoxia-inducible factor-1, HIF-1） and CT-1 （cardiotrophin-1, CT-1）in rat myocardium cell line under hypoxia environment. Meth- ods： The hypoxia environment was achieved by treating cells with cobalt chloride. The viability of H9C2 cells was detected by Cell Counting Kit-8. Chemical synthesized siRNA was transfected into H9C2 ceils by lipofectamine2 000. The mRNA and protein level of HIF-laand CT-1 were detected by real-time PCR and western blot, respectively. Results. After the H9C2 ceils were exposed to cobalt chloride for 48 hours, both the mRNA and protein level of CT-1 in siRNA-HIF-1α transfection group were lower than the siR- NA-control transfection group（P〈0.05）. After the cells were treated with cobalt chloride for three or four days, the viability of H9C2 cells treated with CT-1 was higher thanthe control group （P〈0.05）. The mRNA level of HIF-1 did not change but protein level of HIF-1 increased after exposure of H9C2 cells to CT-I（P〈0.05）. Conclusions： The expression level of HIF-1 decreased with the decrease of CT-1 expres- sion level, and expression level of HIF-1 increased after CT-1 treatment indicating HIF-1 and CT-1 interact with each other in rat myocardium cell line under hypoxia environment.

关 键 词：缺氧诱导因子-1 心肌营养素-1 低氧心肌 

分 类 号：R738[医药卫生—肿瘤]