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
Effects of Sofren on Mitochondrial Function and Dynamics of Hypoxia/Reoxygenation Cardiomyocytes
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摘要: 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。Abstract: 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 optic 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 statistical 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.
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Key words:
- sofren /
- hypoxia/reoxygenation /
- cardiomyocytes /
- ATP /
- ROS /
- MMP /
- OPA1 /
- Drp1 /
- p-DRP1
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