Rg1 prevents myocardial hypoxia/reoxygenation injury by regulating mitochondrial dynamics imbalance via modulation of glutamate dehydrogenase and mitofusin 2
Rg1 prevents myocardial hypoxia/reoxygenation injury by regulating mitochondrial dynamics imbalance via modulation of glutamate dehydrogenase and mitofusin 2
复制标题
Rg1 通过调节谷氨酸脱氢酶和线粒体融合蛋白 2 来调节线粒体动力学失衡，从而预防心肌缺氧/复氧损伤
DOI：
10.1016/j.mito.2015.11.003
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发表时间：
2016-01-01
期刊：
影响因子：
4.4
通讯作者：
Zhou, Hua
中科院分区：
文献类型：
--
作者：
Dong, Gengting;Chen, Tingbo;Zhou, Hua
目的：线粒体功能障碍是缺血性心脏病的一个显著特征，但其潜在的动力学机制(融合/分裂)仍不清楚。在此，我们研究了Rg1在体外缺氧/复氧(H/R)心肌细胞模型中的新功能及其可能的机制。方法：通过MIT、线粒体(活染色)和心脏标志物检测来评估细胞毒性。用ATP含量测定、线粒体内线粒体膜电位测定、ROS、OCR和ECAR法评价线粒体功能。用实时荧光显微镜观察线粒体的动态变化，用高含量图像分析评价线粒体的形态。用Western印迹、RT-PCR和免疫荧光法检测线粒体融合蛋白和分裂相关蛋白的表达。结果：Rg1可抑制GdH的异常调节，并以剂量依赖的方式对抗H/R诱导的细胞损伤和线粒体功能障碍。Rg1显著增加线粒体长度，减少线粒体碎裂的细胞数，上调Mfn2的表达，从而阻止H/R后线粒体动力学的失衡。结论：Rg1通过调节GdH和Mfn2维持线粒体动力学，对H/R诱导的心肌细胞损伤具有保护作用。这些结果为Rg1对心脏I/R障碍的治疗潜力提出了新的保护机制。(C)2015年提交人。Elsevier B.V.和线粒体研究会。
Purpose: Mitochondrial dysfunction is a prominent feature of ischemia heart disease but the underlying mechanism of dynamics (fusion/fission) is still unclear. Here we investigated a novel function and underlying mechanism of Rg1 on an in vitro cardiomyocyte model of hypoxia/reoxygenation (H/R).Methods: Cellular cytotoxicity was evaluated by MIT, mitochondria(viable staining, and cardiac marker detection. Mitochondrial function was evaluated by ATP content measurement, MMP determination, ROS, OCR and ECAR assay. Mitochondrial dynamics was investigated by Live-cell imaging with time-lapse fluorescence microscopy and morphological features were evaluated by the high-content image analysis. Mitochondria(fusion and fission-related proteins, GDH were determined by Western blot RT-PCR and immunofluorescence.Results: Rg1 moderated GDH dysregulation and then protected against H/R-induced cellular damage and mitochondrial dysfunction in a dose-dependent manner. Rg1 significantly increased mitochondrial length, reduced the number of cells with fragmented mitochondria and up-regulated the MFN2 expression finally leading to preventing the imbalance of mitochondrial dynamics following H/R. Knock-down of MFN2 by specific siRNA completely abolished the ability of Rg1 to cell survival by H/R.Conclusion: Rg1 through modulation of GDH and MFN2 maintained mitochondrial dynamics that resulted in protection against H/R-induced cardiomyocyte injury. All these results put forward a new protective mechanism of Rg1 on the therapeutic potential in cardiac I/R disorders. (C) 2015 The Authors. Elsevier B.V. and Mitochondria Research Society.