麝香保心丸对异丙肾上腺素诱导的心肌细胞肥大及其Cx43蛋白表达的影响

Effect of Shexiang Baoxin Pills on isoprenaline-induced myocardial cell hypertrophy and Cx43 expression

目的:观察在麝香保心丸(Shexiang Baoxin Pills,SBP)的影响下异丙肾上腺素(isoprenaline,Iso)诱导的心肌细胞体积、样态及缝隙连接蛋白43(connexin 43,Cx43)表达的变化。方法:采用随机法将H_9C_2心肌细胞分为假手术组(sham组)、Iso组、Iso+SBP组,培养72 h后,在倒置相差显微镜下测定H_9C_2细胞表面积;采用聚氰基丙烯酸正丁酯(bicinchoninic acid,BCA)蛋白检测法测定细胞蛋白浓度;采用MTT比色法测定心肌细胞存活率;采用Western印迹检测Cx43蛋白的表达。结果:倒置相差显微镜下观察发现:与sham组比较,Iso组心肌细胞表面积和细胞蛋白浓度均增加(均P<0.05);而与Iso组相比,Iso+SBP组心肌细胞表面积减少,细胞蛋白浓度降低(均P<0.05);Western印迹显示:与sham组相比,Iso组H_9C_2细胞Cx43蛋白的表达水平明显下调(P<0.05);与Iso组相比,Iso+SBP组Cx43的表达水平明显上调(P<0.05)。结论:麝香保心丸可改善Iso诱导的心肌细胞肥大,并能使Cx43蛋白表达下调。

Objective： To observe the effects of Shexiang Baoxin Pill（SBP） on isoprenaline（Iso）-induced changes in myocardial cell volume, shape, and connexin 43（Cx43） expression. Methods： H9C2 myocardial cells were randomly divided into a control group, a Iso group and a Iso＋SBP group. After 72 h of culture, the average surface area of H9C2 cells was measured under phase contrast microscope. Bicinchoninic acid（BCA） protein assay was carried out to determine the concentration of proteins. The survival rate of myocardial cells was measured by methylthiazolyldiphenyl-tetrazolium bromide（MTT） assay, and the Cx43 expression was detected by Western blot. Results： The mean surface area and Cx43 concentration in Iso-treated myocardial cells were increased under the phase contrast microscope（P〈0.05）. Compared with the Iso group, the mean surface area was decreased, and the Cx43 concentration was reduced in the Iso＋SBP group（both P〈0.05）. Compared with the control group, the Cx43 expression was obviously down-regulated in the H9C2 cells of the Iso group（P〈0.05）; while compared with the Iso group, the Cx43 expression was obviously up-regulated in the Iso＋SBP group（P〈0.05）. Conclusion： Shexiang Baoxin Pills can prevent Iso-induced myocardial hypertrophy and downregulate Cx43 expression.