Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University , Hangzhou 310058, P.R. China.
College of Preclinical Medicine, Zhejiang Chinese Medical University , Hangzhou 310053, P.R. China.
J Proteome Res. 2018 Feb 2;17(2):794-803. doi: 10.1021/acs.jproteome.7b00450. Epub 2018 Jan 23.
Shenmai formula (SM) has been a traditional medicinal remedy for treating cardiovascular diseases in China for 800 years; however, its mechanism of action remains unclear. To explore the mechanism underlying cardioprotective effects of SM, iTRAQ-based proteomic approach was applied to analyze protein of myocardium in rats with myocardial ischemic injury. Upon treatment with SM and its two major components Red ginseng (RG) and Radix Ophiopogonis (OP), 101 differentially expressed proteins were filtered from a total of 712 detected and annotated proteins. They can be classified according to their locations and functions, while most of them are located in intracellular organelle, participating in cellular metabolic process. The functions of them are mostly associated with mitochondrial oxidative phosphorylation/respiration. The differentially expressed proteins were validated by liquid chromatography-tandem mass spectrometry and Western blotting (ATP5D, NDUFB10, TNNC1). Further in vitro experiments found that SM could attenuate hypoxia induced impairment of mitochondrial membrane potential and cellular ATP concentration in neonatal rat ventricular myocytes. Interestingly, the result of quantitative mitochondrial biogenesis assays revealed that SM had dominant positive effects on the maximum respiration, ATP-coupled respiration, and spare capacity of mitochondria in response to hypoxia. Hence, our findings suggest that SM promotes mitochondrial function to protect cardiomyocytes against hypoxia, which provides a possible illustration for conventional botanical therapy on a molecular level.
参麦方（SM）是中国治疗心血管疾病的传统药物已有 800 年的历史，但作用机制尚不清楚。为了探讨 SM 对心肌缺血损伤的心脏保护作用机制，我们采用 iTRAQ 基于蛋白质组学的方法分析了心肌缺血损伤大鼠心肌中的蛋白质。在用 SM 及其两个主要成分红参（RG）和麦冬（OP）进行治疗后，从总共检测到的 712 个注释蛋白中筛选出 101 个差异表达蛋白。它们可以根据其位置和功能进行分类，其中大多数位于细胞内细胞器中，参与细胞代谢过程。它们的功能主要与线粒体氧化磷酸化/呼吸有关。通过液相色谱-串联质谱法和 Western blot 验证了差异表达蛋白（ATP5D、NDUFB10、TNNC1）。进一步的体外实验发现，SM 可以减轻缺氧诱导的乳鼠心室肌细胞线粒体膜电位和细胞 ATP 浓度的损伤。有趣的是，定量线粒体生物发生测定的结果表明，SM 对缺氧时线粒体的最大呼吸、ATP 偶联呼吸和备用能力具有明显的正向作用。因此，我们的研究结果表明，SM 通过促进线粒体功能来保护心肌细胞免受缺氧损伤，这为传统植物疗法提供了一个分子水平上的可能解释。
