摘要
目的检测缝隙连接通道蛋白43(Conexin43,CX43)在缺血再灌注中的心肌细胞基因表达和CX43通道蛋白变化情况,分析CX43变化的原因,判断CX43在心肌缺血再灌注中所起得作用。方法原代培养心肌细胞,建立缺血再灌注模型,分别于正常、缺氧30 min、再灌注1h、2 h、3 h、6 h搜集细胞。逆转录聚合酶链式反应(RT-PCR)检测CX43mRNA表达、蛋白免役印迹法(Western blot)检测CX43蛋白的含量。结果未用药干预的心肌细胞缺氧30 min时CX43mRNA表达与蛋白含量和正常相比无显著差异(P＞0．05),再供氧1h、2 h、3 h、6 h则分别减少了39．16%、45．00%、46．67%、51．67%。和正常相比差异显著(P＜0．01)。其中再供氧1 h下降幅度最大。结论CX43在心肌细胞缺氧再灌注过程中mRNA表达与蛋白含量均是减低的,其中再供氧1 h下降幅度最大。
Objective To investigate the expression of conexin43 （ CX43 ） gene and CX43 protein in myocardial cells iscbemic reperfusion. The relation between the expression of CX43 and myocardial cell ischemia was studied. Method A culture system of neonatal cardiac myocytes and the model of the myocardial cells iscbemic reperfusion were establisbed, The cells were collected at normal, ischemia for 30min, reperfusion for 1 h, 2 h, 3 h, 6 h. The mRNA level of cells was determined with reverse transcriptase polymerase chain reaction（RT-PCR）. The content of protein was detected by western blotting. Results In control, there are no significantly different between the CX43 expression of myocytes iscbemia for 30rain and normal（P 〉 0.05）. There were decreases in myocytes reperfusion for 1 h,2 h,3 h,6 h（P 〈 0.01 ）. Conclusions The CX43 mRNA level of cells and the content of CX43 protein significantly decrease during myocytes reperfusion. Of which, the range of decrease is the largest in reperfusion for 1 h.
出处
《中国分子心脏病学杂志》
CAS
2007年第1期27-29,共3页
Molecular Cardiology of China
