Objective: To investigate the effect of salvianolic acid A (Sal A) on mitochondrial autophagy and cell injury in human pulmonary microvascular endothelial cells (HPMVECs) injury induced by lipopolysac-charide (LPS). Methods: LPS 10 μg/mL was used to establish cell injury in vitro. The cells were divided into four groups: control group, Sal A group, LPS group, LPS+Sal A group. MTT was used to analyze cell proliferation. The reactive oxygen species (ROS) content and mitochondrial membrane potential were detected by flow cytom-etry. Western blot was used to analyze the expression of microtubules associated protein light chain 3 (LC3)-II/I, Beclin1, PTEN mediated putative kinase protein 1 (PINK1) and Parkin. Results: LPS decreased the cell viability in time-dependent manner. The viability in LPS+Sal A group was higher than the LPS group, but lower than the control group (P<0.05). Compared with the control group (100%), the ROS content (283.96%±10.88%) increased and the level of membrane (38.25%±5.69%) potential decreased in LPS group (P<0.05). Compared with LPS group, ROS content (138.76%±11.82%) decreased, and the mitochondrial membrane potential (76.56%±6.22%) increased in LPS+Sal A group (P<0.05). The expression of LC-3II/I, Beclin1, PINK1 and Parkin in LPS group was higher than the control group. The expression of LC-3II/I, Beclin1, PINK1 and Parkin in LPS+Sal A group was lower than LPS group. Conclusion: Sal A alleviates the proliferation inhibition by LPS, which may be re-lated with decreased intracellular ROS content, stabilization of mitochondrial membrane potential and mitochon-drial autophagy.%目的:探讨丹酚酸A(Sal A)对脂多糖(LPS)诱导的人肺微血管内皮细胞(HPMVECs)线粒体自噬和细胞损伤的影响.方法:采用终浓度为10 μg/mL LPS构建细胞损伤模型,并将实验分为对照组、Sal A组、LPS组、LPS+Sal A组.采用噻唑蓝染色法分析细胞增殖情况,流式细胞技术检测细胞内活性氧(ROS)含量和线粒体膜电位水平,蛋白免疫印迹法检测微管相关蛋白轻链3(LC3)-II/I、Beclin1、PTEN介导的假定激酶蛋白1(PINK1)和Parkin蛋白相对表达量.结果:10 μg/mL LPS呈时间依赖性抑制HPMVECs的增殖;LPS+Sal A组细胞相对存活率高于LPS组,低于对照组(P<0.05).LPS组ROS含量(283.96%±10.88%)增高,膜电位水平(38.25%±5.69%)降低,与对照组(100%)相比差异有统计学意义(P<0.05);LPS+Sal A组ROS含量(138.76%±11.82%)低于LPS组,线粒体膜电位水平(76.56%±6.22%)高于LPS组(P<0.05).LPS组细胞LC3-II/I、Beclin1、PINK1和Parkin蛋白表达量较对照组增高(P<0.05),LPS+Sal A组LC3-II/I、Beclin1、PINK1和Parkin蛋白相对表达量低于LPS组(P<0.05);LPS组线粒体PINK1和Parkin蛋白表达量与对照组比较增高(P<0.05),LPS+Sal A组明显低于LPS组(P<0.05).结论:Sal A减轻LPS诱导的HPMVECs细胞细胞损伤作用,其可能机制与其降低细胞内ROS,稳定线粒体膜电位,抑制线粒体自噬有关.
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