α-硫辛酸对H9c2心肌细胞损伤保护作用的研究

Protective effect of α-lipoic acid on myocardial injury of H9c2 cells

目的研究α-硫辛酸(ALA)对H9c2心肌细胞损伤的保护作用。方法建立H9c2心肌细胞的缺氧复氧模型,将H9c2心肌细胞为4组。①正常对照组,不做干预。②ALA组:在细胞培养基中加入ALA。③H/R组:对心肌细胞采用缺氧复氧干预。④ALA+H/R组:同时采用ALA和缺氧复氧干预。通过显微镜观察细胞状态,采用MTT检测细胞活性及流式细胞仪检测细胞凋亡,采用JC-1检测线粒体膜电位。结果①ALA+H/R组细胞死亡数量较H/R组H9c2细胞死亡明显减少,可见有细胞呈扁平梭形,也可见细胞贴壁率、折光性下降,细胞中颗粒增多,连接松散,细胞间可见较多细胞碎片。②H/R组缺氧复氧诱导后细胞活性(62.5%)较对照组(100%)与ALA组(98.6%)明显降低,差异有统计学意义(均P<0.05);ALA+H/R组处理后细胞活性(83.7%)较H/R组明显升高,差异有统计学意义(P<0.05)。③H/R组缺氧复氧诱导后细胞凋亡比例(45.8%)较对照组(9.8%)与ALA组(10.7%)明显增加,差异有统计学意义(均P<0.05);ALA+H/R组处理后细胞凋亡比例(24.5%)较H/R组明显减少,差异有统计学意义(P<0.05)。④H/R+ALA组线粒体膜电位为(9.90±0.19)V,H/R组为(6.90±0.27)V,差异有统计学意义(P<0.05)。结论ALA对缺氧复氧诱导H9c2细胞损伤起保护作用。

Objective To investigate the protective effects of α-lipoic acid(ALA) on H9c2 cells exposed to hypoxia/reoxygenation. Methods H9c2 myocardial cell model was established, H9c2 cells were assigned to 4 groups.①Control group.②ALA group (ALA): ALA was added to cell culture medium.③H/R group (H/R): Cardiomyocytes were treated with anoxic reoxygenation.④ALA+H/R group (ALA+H/R): ALA and anoxic reoxygenation were used simultaneously. The cell state was observed by microscope, the cell activity was detected by MTT, the apoptosis was detected by flow cytometry, and the mitochondrial membrane potential was measured by JC-1. Results ①In ALA+H/R group, the number of cell death was significantly reduced than that in H/R group, It can be seen that some cells were flat and spindle-shaped, but also that the cell adherence rate and refraction were decreased, the particles in the cells were increased and the connections are loose, and there were many cell fragments between the cells.②After hypoxia reoxygenation, the cell activity in H/R group (62.5%) were significantly lower than that in control group (100%) and ALA group (98.6%), and the differences were statistically significant (all P < 0.05).③Compared with the control group (9.8%) and ALA group (10.7%), the proportion of apoptosis in H/R group (45.8%) after hypoxia reoxygenation were significantly increased (all P < 0.05). Apoptosis rate (24.5%) in ALA+H/R group was significantly lower than that in H/R group ( P < 0.05).④The mitochondrial membrane potential of the H/R+ALA group was (9.90±0.19) V, and that of the H/R group was (6.90±0.27) V, and the difference was statistically significant ( P < 0.05). Conclusion ALA has a protective effect on H9c2 cell injury induced by hypoxia reoxygenation.