大株红景天对缺氧/复氧大鼠原代心肌细胞线粒体功能及线粒体动力学的影响

摘要

目的 通过观察大株红景天对缺氧/复氧(Hypoxia/reoxygenation,H/R)心肌细胞线粒体功能及线粒体动力学的影响,探讨其抗心肌细胞 H/R损伤的可能机制.方法 分离并培养新生 SD大鼠原代心肌细胞,选符合实验要求的心肌细胞,随机分为5组:空白对照组、缺氧/复氧组(H/R组)、红景天前干预组(S+H/R组)、红景天后干预组(H/R+S组)、红景天前后干预组(S+H/R+S组).实验结束后多功能酶标仪测定心肌细胞内三磷酸腺苷(ATP)水平、活性氧簇(ROS)含量,JC-1法测定心肌细胞线粒体膜电位(MMP),Western blot 法检测线粒体动力学相关蛋白视神经萎缩蛋白(OPA1)、动力相关蛋白1 (DRP1)及磷酸化动力相关蛋白1(p-DRP1)的表达水平.结果 S+H/R组、H/R+S组及 S+H/R+S组 H/R 心肌细胞内ATP含量均增加,ROS水平降低,并抑制 MMP的降低,上调 OPA1蛋白表达,抑制 p-Drp1蛋白表达(P<0.05);S+H/R+S组分别与 S+H/R组、H/R+S组组间比较,ATP、ROS、MMP 水平变化及 p-Drp1表达量均具有统计学差异(P<0.05),而OPA1蛋白的表达无明显差异;S+H/R组、H/R+S组及 S+H/R+S组组间比较,Drp1的表达量无明显差异.结论 大株红景天对 H/R损伤的心肌细胞有保护作用,其机制可能与改善受损心肌细胞线粒体功能障碍、稳定线粒体动力学平衡有关,值得进一步探讨.%OBJECTIVE To explore the effects of Sofren on mitochondrial dysfunction and dynamic unbalance induced by H/R injury.METHODS Primary neonatal rat cardiomyocytes were cultured and randomly assigned to five groups based on the sofren that was used and the treatment simulating reperfusion injury:control group,H/R group,sofren preconditioning+H/R group(S+H/R),H/R+sofren postconditioning group(H/R+S),sofren pre-and postconditioning +H/R group(S+H/R+S).The intracellular adenosine triphosphate(ATP)level,reactive oxygen species(ROS)accumulation was measured by multifunctional microplate reader,mitochondrial membrane potential(MMP)was measured though JC-1,mitochondrial op-tic atrophy(OPA1),dynamin-related protein 1(Drp1)and phosphorylation dynamin-related 1(p-Drp1)was measured through Western blot in each group.RESULTS The ROS accumulation was inhibited,MMP and higher ATP level was maintained, higher OPA1 and lower p-DRP1 was expressed,preconditioning or postconditioning or pre-and postconditioning by sofren(P<0.05);there is statistical difference of ATP,ROS and MMP values and p-DRP1 expression between S+H/R+S group and S+H/R group,as well as S+H/R+S group and H/R+S,while no statistical difference of OPA1 expression;there is no sta-tistical difference of DRP1 expression between all the five groups.CONCLUSION These results showed that sofren exerted a protective effect on cardiomyocytes injuried by H/R,and the machanism may correlate with rescuing H/R-induced myocardial mitochondrial dysfunction and dynamic unbalance,and further studies are required.