Abstract Context Simvastatin is the first line therapeutic drug for coronary heart disease and atherosclerosis. The protective effect mechanism of simvastatin on cardiomyocytes is unclear. Objective This study explores the effect of simvastatin on high glucose induced cardiomyocyte injury and the role of autophagy during the process. Materials and methods H9c2 cells were incubated with different doses of glucose (0, 50, 100, 200 mM) for 24 h to verify the glucose induced injury. The H9c2 cells were pre-treated with simvastatin at different dosages (0, 0.1, 0.5, 1 μM) for 30 min to rescue the injury followed by the autophagy evaluation. 3-MA was used as an autophagy inhibitor to confirm the role of autophagy in simvastatin treated process. CCK-8 assay, FACS assay, confocal microscopy, western blotting and immunofluorescence analysis were conducted to evaluate the high glucose induced injury or protective effects of simvastatin in H9c2 cell line. Results High glucose dramatically decreased H9c2 cell viability (0 mM, 0.58 ± 0.09%; vs. 50 mM, 8.67 ± 0.43%; 100 mM, 16.1 ± 3.56%; 200 mM, 32.9 ± 2.63%), induced significant cell apoptosis (0 mM, 0.96 ± 0.16%, vs. 50 mM, 7.00 ± 0.63%; 100 mM, 12.9 ± 0.78%; 200 mM, 21.8 ± 1.17%) and suppressed cell autophagy. Simvastatin decreased apoptosis and attenuate injury by decreasing cell apoptosis ratio, elevating Bcl-2 expression while decreasing Bax and caspase-3 protein expressions. Meanwhile, simvastatin restored the autophagy depicted by western blotting with increased ATG-5, Beclin1 and LC3II/LC3I protein expression and decreased p62 expression, as well as immunofluorescence with elevated LC3 fluorescence density. Discussion and conclusions The myocardial protective effect mediated by autophagy activated by simvastatin to some extent elucidated the mechanism of the protective effect of simvastatin on H9c2 cell injury, which provided a certain theoretical basis for the clinical application of simvastatin in the treatment of cardiovascular diseases. In addition, we speculate that simvastatin may be used for diabetes associated cardiovascular diseases.

中文翻译：

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辛伐他汀通过诱导自噬保护高糖诱导的 H9c2 细胞免受损伤

摘要 背景 辛伐他汀是冠心病和动脉粥样硬化的一线治疗药物。辛伐他汀对心肌细胞的保护作用机制尚不清楚。目的本研究探讨辛伐他汀对高糖诱导心肌细胞损伤的影响及自噬在此过程中的作用。材料和方法 H9c2 细胞与不同剂量的葡萄糖（0、50、100、200 mM）孵育 24 小时以验证葡萄糖诱导的损伤。H9c2 细胞用不同剂量（0、0.1、0.5、1 μM）的辛伐他汀预处理 30 分钟以挽救损伤，然后进行自噬评估。3-MA 被用作自噬抑制剂以证实自噬在辛伐他汀处理过程中的作用。CCK-8 测定、FACS 测定、共聚焦显微镜、进行蛋白质印迹和免疫荧光分析以评估高糖诱导的辛伐他汀对H9c2细胞系的损伤或保护作用。结果 高糖显着降低 H9c2 细胞活力（0 mM，0.58 ± 0.09%；对比 50 mM，8.67 ± 0.43%；100 mM，16.1 ± 3.56%；200 mM，32.9 ± 2.63%），诱导显着的细胞凋亡（0 mM, 0.96 ± 0.16%, vs. 50 mM, 7.00 ± 0.63%; 100 mM, 12.9 ± 0.78%; 200 mM, 21.8 ± 1.17%) 并抑制细胞自噬。辛伐他汀通过降低细胞凋亡率、提高 Bcl-2 表达同时降低 Bax 和 caspase-3 蛋白表达来减少细胞凋亡并减轻损伤。同时，辛伐他汀恢复了蛋白质印迹描述的自噬，增加了 ATG-5、Beclin1 和 LC3II/LC3I 蛋白表达，降低了 p62 表达，以及具有升高的 LC3 荧光密度的免疫荧光。讨论与结论 辛伐他汀激活自噬介导的心肌保护作用在一定程度上阐明了辛伐他汀对H9c2细胞损伤保护作用的机制，为辛伐他汀治疗心血管疾病的临床应用提供了一定的理论依据。此外，我们推测辛伐他汀可能用于糖尿病相关的心血管疾病。为辛伐他汀临床应用治疗心血管疾病提供了一定的理论依据。此外，我们推测辛伐他汀可能用于糖尿病相关的心血管疾病。为辛伐他汀临床应用治疗心血管疾病提供了一定的理论依据。此外，我们推测辛伐他汀可能用于糖尿病相关的心血管疾病。