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目的:检测卡维地洛(Cavedilol,CVD)对缺氧再供氧时心肌细胞缝隙连接通道蛋白43(Conexin43,CX43)的基因表达和CX43通道蛋白变化的影响。方法:原代培养心肌细胞,随机分为2组:未用药组,CVD组。建立缺氧、再供氧模型,分别于正常、缺氧30min、再供氧1h、2h、3h、6h搜集细胞。逆转录聚合酶链式反应(RT-PCR)检测CX43 mRNA表达、蛋白免役印迹法(Western blot)检测CX43蛋白的含量。结果:未用药组的心肌细胞缺氧30min时CX43 mRNA表达与蛋白含量和正常相比无显著差异(P>0.05)。再供氧1h、2h、3h、6h则分别减少了39.16%、45.00%、46.67%、51.67%,和正常时相比差异显著(P<0.01)。其中再供氧1h下降幅度最大。用CVD干预后再供氧1h、2h、3h、6h则分别减少了22.95%、27.87%、30.33%、35.25%(P<0.01)。再供氧1h、6h的下降幅较未用药组分别减少了41.39%、31.78%,差异显著(P<0.01)。结论:心肌细胞缺氧再供氧时,其CX43基因表达和蛋白含量明显减少,卡维地洛可抑制CX43降解。
AIM: To investigate the effect of cavedilol (CVD) on the gene expression of CX43 and the CX43 channel in hypoxia and reoxygenation. Methods: Primary cultured cardiomyocytes were randomly divided into two groups: non-medication group and CVD group. Establish hypoxia, reoxygenation model, respectively, in normal, hypoxia 30min, then oxygen 1h, 2h, 3h, 6h cells were collected. The expression of CX43 mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and the content of CX43 protein was detected by Western blot. Results: There was no significant difference in CX43 mRNA expression between the untreated cardiomyocytes and the normal group at 30min hypoxia (P> 0.05). After reoxygenation for 1h, 2h, 3h, 6h, they decreased by 39.16%, 45.00%, 46.67%, 51.67%, respectively, which was significantly different from that of the normal one (P <0.01). Which 1h oxygen maximum decline. After intervention with CVD for 1h, 2h, 3h, 6h, they decreased by 22.95%, 27.87%, 30.33%, 35.25% respectively (P <0.01). Compared with the control group, the decreasing rate of reoxygenation for 1h and 6h decreased by 41.39% and 31.78% respectively (P <0.01). CONCLUSION: When hypoxia and reoxygenation of cardiomyocytes, the CX43 gene expression and protein content are significantly reduced. Carvedilol can inhibit the CX43 degradation.