在模拟低氧环境下心肌细胞HIF-1与CT-1的相互作用

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。

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

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.Methods: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 cells by lipofectamine2 000.The mRNA and protein level of HIF-1αand CT-1 were detected by real-time PCR and western blot,respectively.Results:After the H9C2 cells 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 siRNA-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-1(P<0.05).Conclusions:The expression level of HIF-1 decreased with the decrease of CT-1 expression 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.