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目的:从钙调素蛋白依赖激酶Ⅱ(Ca MKⅡ)信号通路的角度探讨黄芪甲苷(AsⅣ)对异丙肾上腺素(Iso)诱导乳鼠心肌细胞肥大的保护作用及其可能机制。方法:原代培养新生乳鼠心肌细胞,10 mol/L异丙肾上腺素诱导心肌细胞肥大,观察KN93(Ca MKⅡ抑制剂)、黄芪甲苷3,10,30 mol/L剂量组对肥大细胞的影响。采用消化分离法及计算机图像分析系统检测心肌细胞体积;考马斯亮蓝试剂盒检测心肌细胞中总蛋白含量;罗丹明-鬼笔环肽染色观察细胞骨架;反转录聚合酶链反应(RT-PCR)法检测心房利钠肽(ANP)mRNA表达;Wertern blot检测心肌细胞Ca MKⅡ表达。结果:与正常对照组细胞比较,异丙肾上腺素模型组细胞体积、总蛋白含量、ANPmRNA和Ca MKⅡ表达分别增加98.14%、52.63%、87.37%和55.22%。黄芪甲苷可有效抑制异丙肾上腺素诱导的心肌细胞肥大,黄芪甲苷30 mol/L组可抑制异丙肾上腺素诱导的心肌肥大,使模型组细胞体积减小49.67%、总蛋白含量降低33.87%、ANPmRNA表达降低40.00%,Ca MKⅡ的表达降低31.73%。结论:黄芪甲苷可抑制异丙肾上腺素诱导的心肌细胞肥大,其机制可能与抑制Ca MKⅡ信号通路有关。
OBJECTIVE: To investigate the protective effects of AsⅣ on cardiomyocyte hypertrophy induced by isoprenaline (Iso) and its possible mechanism from the perspective of Ca ~ (2 +) signaling pathway. Methods: Primary cultured neonatal rat cardiomyocytes were induced by 10 mol / L isoprenaline to induce cardiomyocyte hypertrophy. The effects of KN93 (Ca MK Ⅱ inhibitor), astragaloside 3,10 and 30 mol / L on mast cells influences. Cardiomyocyte volume was detected by digestion and computerized image analysis system; Coomassie Brilliant Blue Kit was used to detect total protein in cardiomyocytes; Rhodamine-phalloidin staining was used to observe the cytoskeleton; RT-PCR ) Method was used to detect the mRNA expression of atrial natriuretic peptide (ANP). The expression of Ca MKⅡ in cardiomyocytes was detected by Wertern blot. Results: Compared with the normal control group, the cell volume, total protein content, ANP mRNA and Ca MKⅡ expression increased by 98.14%, 52.63%, 87.37% and 55.22% in the isoproterenol model group. Astragaloside IV could effectively inhibit isoprenaline-induced cardiomyocyte hypertrophy. Astragaloside 30 mol / L inhibited isoprenaline-induced cardiac hypertrophy, decreased the volume of the model group by 49.67% and decreased the total protein content by 33.87 %, The expression of ANP mRNA decreased by 40.00% and the expression of CaMKII decreased by 31.73%. Conclusion: Astragaloside IV can inhibit isoprenaline-induced cardiomyocyte hypertrophy, and its mechanism may be related to the inhibition of Ca MK Ⅱ signaling pathway.