红景天苷对H9c2心肌细胞应激高糖损伤的保护作用及机制研究
Protective Effect and Mechanism of Salidroside on H9c2 Cardiomyocytes Under Oxidative Stress and Hyperglycemia
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摘要: 目的 利用氧化应激联合高糖复合损伤心肌细胞，建立H9c2心肌细胞应激及高糖损伤模型，探讨应激高糖H9c2心肌细胞线粒体动力学相关蛋白表达、线粒体膜电位水平及细胞凋亡的影响，以及红景天苷对应激高糖H9c2心肌细胞的保护作用及其机制。方法 将正常培养的H9c2细胞随机分为5组：①对照组（C组，正常培养基）；②高糖组（G组，高糖培养基）；③H2O2组（H组，H2O2处理2小时后，更换为正常培养基继续培养）；④高糖+H2O2组（GH组，H2O2及高糖处理2小时后，更换为高糖培养基继续培养）；⑤红景天苷+高糖+H2O2组（SGH组，H2O2+高糖+红景天苷处理2小时后，更换为红景天苷+高糖培养基继续培养）。分组加药培养后，利用流式细胞技术分析各组H9c2心肌细胞的凋亡率；Western blot分别检测线粒体动力学相关蛋白Drp1、OPA1的表达水平；JC-1荧光染色法检测各组心肌细胞的线粒体膜电位水平。结果 ①与C组比较，G组H9c2细胞Drp1蛋白表达水平显著升高，OPA1蛋白表达水平显著降低（P<0.01）；线粒体膜电位下降，细胞凋亡率升高（P＜0.05）；与C组比较，H组及GH组H9c2细胞Drp1蛋白表达水平显著升高，OPA1蛋白表达水平显著降低，线粒体膜电位显著下降，细胞凋亡率显著升高（P<0.01）。②与GH组比较，SGH组Drp1蛋白表达水平显著降低，OPA1蛋白表达水平显著升高，线粒体膜电位显著升高，细胞凋亡率显著降低（P<0.01）。结论 ①氧化应激联合高糖通过降低线粒体膜电位，抑制H9c2心肌细胞线粒体动力学平衡，增加心肌细胞凋亡，从而损伤心肌细胞。②红景天苷可抑制氧化应激联合高糖引起的H9c2心肌细胞凋亡，其机制与升高线粒体膜电位水平以及增加线粒体OPA1蛋白表达、抑制Drp1蛋白表达从而改善线粒体动力学平衡有关；红景天苷对应激高糖心肌细胞具有一定的保护作用，值得进一步深入研究。Abstract: OBJECTIVE To investigate the protective effects of Salidroside in H9c2 cardiomyocytes under oxidative stress and hyperglycemia. METHODS H9c2 cardiomyocytes were divided into five groups： ①the control group （the cells treated with normal glucose）；②the high glucose group （the cells treated with high glucose）； ③the H2O2 group （the cells treated with H2O2 for 2 hours）； ④the high glucose+H2O2 group （the cells treated with H2O2 and high glucose）； ⑤the high glucose+H2O2+Salidroside group（the cells treated with H2O2 and high glucose and salidroside）. Activity of H9c2 cardiomyocytes was detected by cell counting kit （CCK-8）.The rate of apoptotic cells were detected by flow cytometry.The expression of OPA1 and Drp1 protein in mitochondria was determined by Western blot.Mitochondria membrane potentials were measured by JC-1 kit. RESULTS ①Compared with the control groups，the expression of OPA1 cells decreased （P<0.01），the expression of Drp1 cells increased （P<0.01），accompanied by reduction in mitochondrial membrane potential and the rate of apoptotic cells were increased （P<0.05）in high glucose group；Compared with the control groups，the expression of OPA1 cells decreased （P<0.01），the expression of Drp1 cells increased（P<0.01），accompanied by reduction in mitochondrial membrane potential and the rate of apoptotic cells were increased（P<0.01）in H2O2 group and high glucose+ H2O2 group；②Compared with the high glucose+H2O2 group，the expression of OPA1 cells increased（P<0.01），the expression of Drp1 cells decreased（P<0.01），accompanied by increased mitochondrial membrane potential and the rate of apoptotic cells were decreased （P < 0.01） in salidroside+ high glucose+ H2O2 group. CONCLUSION ①Oxidative stress and hyperglycemia can inhibit the balance of mitochondrial dynamics of H9c2 cardiomyocytes， reduce the mitochondrial membrane potential， and increase the apoptosis of cardiomyocytes， so as to damage cardiomyocytes. ②Salidroside can alleviate injury under oxidative stress and hyperglycemia，reduce cardiomyocyte apoptosis and protect cell mitochondrial function， and its mechanism is related to increased expression of OPA1 and decreased expression of Drp1，to improve the mitochondrial dynamic balance， and the increase of mitochondrial membrane potential level. Salidroside has a protective effect on oxidative stress and hyperglycemia， which deserves further study.
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Key words:
- H9c2 cardiomyocytes /
- salidroside /
- oxidative stress /
- hyperglycemia /
- mitochondrial dynamics /
- apoptosis
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