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目的:探究丹参主要成分丹参酮ⅡA能否减轻心肌细胞缺氧/复氧(H/R)损伤,以及对其诱导的自噬相关机制的研究。方法:在乳鼠心肌细胞原代培养的基础上,构建缺氧6h复氧18h的细胞损伤模型。应用丹参酮ⅡA预处理,检测细胞活力、m RFP-GFP-LC3双荧光标记的自噬潮的变化及自噬相关蛋白P62、LC3Ⅱ、Beclin-1及Bcl-2的表达。结果:H/R处理引起细胞损伤甚至死亡。给予丹参酮ⅡA预处理后,可以显著改善心肌细胞活性,抑制自噬潮,并降低由H/R引起的自噬相关蛋白P62、LC3Ⅱ及Beclin-1的表达,同时增强抗凋亡蛋白Bcl-2的表达。在加入100nmol/L的Rapamycin处理后,上述蛋白的表达受到抑制。结论:丹参酮ⅡA可能是通过激活m TOR信号通路抑制过度自噬来减轻心肌细胞缺氧/复氧损伤的。
Objective: To investigate whether Tanshinone ⅡA, the main component of Radix Salviae Miltiorrhizae, can reduce the hypoxia / reoxygenation (H / R) injury of cardiomyocytes and the related mechanism of autophagy induced by Tanshinone ⅡA. Methods: On the basis of primary culture of neonatal rat cardiomyocytes, a cell injury model of hypoxia for 6 hours and reoxygenation for 18 hours was constructed. TanshinoneⅡA pretreatment was used to detect the cell viability, the changes of m RFP-GFP-LC3 double fluorescently labeled autophagy and the expression of autophagy-related proteins P62, LC3Ⅱ, Beclin-1 and Bcl-2. Results: H / R treatment caused cell damage or even death. Pretreatment with tanshinone Ⅱ A could significantly improve the activity of cardiomyocytes, inhibit the autophagy and reduce the expression of autophagy-related proteins P62, LC3Ⅱ and Beclin-1 induced by H / R, and enhance the anti-apoptotic protein Bcl-2 expression. The expression of the above protein was inhibited after treatment with 100 nmol / L Rapamycin. Conclusion: Tanshinone ⅡA may reduce myocardial hypoxia / reoxygenation injury by activating m TOR signaling pathway to inhibit over-autophagy.