淫羊藿苷改善高糖培养乳鼠心肌细胞活性的线粒体相关机制

Icariin improves the viability of the neonatal rat cardiomyocyte via mitochondria in high Glucose

目的研究淫羊藿苷对抗体外高浓度葡萄糖诱导乳鼠心肌细胞的损伤及其潜在的线粒体相关机制。方法SD乳鼠心肌细胞分别在含5.5mmol／L葡萄糖（正常对照组）、5．5mmol／L葡萄糖＋20.0mmol／L甘露醇（高渗对照组）、25.5mmol／L葡萄糖（高糖损伤组）、25.5mmol／L葡萄糖＋10μmol／L淫羊藿苷（低剂量淫羊藿昔保护组）和25.5mmol／L葡萄糖＋100μmol／L淫羊藿苷（高剂量淫羊藿苷保护组）的培养基中进行原代培养48h后，检测细胞活力、细胞培养上清液乳酸脱氢酶（LDH）含量、细胞活性氧自由基（ROS）生成、超氧化物歧化酶（SOD）活性，同时检测心肌线粒体跨膜电位，并检测线粒体通透性转换孔（MPTP）开放值。结果与正常对照组比较，高糖培养组心肌细胞活性、细胞SOD活性及心肌线粒体跨膜电位水平均显著下降（P〈0．01），而培养上清液LDH含量、细胞ROS生成及心肌细胞MPTP开放值则明显升高（P〈001）。淫羊藿苷干预减轻了高糖对心肌细胞各项指标的损伤，尤其是高剂量淫羊藿苷保护组的细胞活性、LDH漏出量、ROS生成值、SOD活性、MPTP开放值、线粒体跨膜电位均有明显改善。高渗对照组各项指标与正常对照组相比差异无统计学意义。结论淫羊藿苷对体外高糖培养乳鼠心肌细胞具有保护作用，其作用机制可能通过提高细月旬抗眚化能力．减少ROS诱导的心肌MPTP开放程度．维持线粒体正常跨膜电位而实现的。

Objective To investigate the protective effect of icariin against the injury of neonatal rat cardiomyocytes induced by high glucose and its underlying mitochondrial mechanisms. Methods The primitive cardiomyocytes was respectively cultured with 5.5 mmol/L glucose（Control group）, 5.5 mmol/L Glu ＋ 20.0 mmol/L mannitol （High osmotic group, HO）, 25.5 mmol/L Glu （High glucose group, HG）, 25.5 mmol/L Glu ＋ 10 v mol/L icariin （Low dose icariin group, LICA）, 25.5 mmol/L Glu ＋100 μmol/L icariin （High dose icariin group, HICA） for 48 h. The cell viability, the release of lactate dehydrogenase （LDH） from cells, the reac- tive oxygen species （ROS） and the activity of superoxide dismutase （SOD） were measured. The mitochondrial membrane potential （△ψm） and the mitochondrial permeability transition pore were assayed. Results The cell viability, activity of SOD, and the △ψm were markedly decreased （P〈0.01）, whereas, the leakage of cell LDH, ROS level, and the opening of MPTP were significantly increased in HG group （P〈0.01）. Treatment with ICA reduced the cardiomyocyte injury induced by high glucose, especially in the HICA group, the ceil viability, the leakage of LDH, ROS level, the activity of SOD, the opening of MPTP, and the△ψm were significantly improved compared with those in HG group. There was no significant difference in all parameters between CG group and HO group. Conclusion The icariin protected cardiomyocytes against the injury induced by high glucose in vitro, and the mechanism may be associated with enhancing the antioxidative ability, reducing the opening of MPTP induced by ROS, and maintaining the normal mitochondrial membrane potential.