[Proteomic and metabolomic profiling of the rat model of myocardial ischemia-reperfusion injury treated with electroacupuncture].

摘要

This study aims to explore the protective effect of electroacupuncture (EA) on myocardial ischemia-reperfusion injury (MIRI) and decipher the molecular mechanism. Forty male SD rats were randomly allocated into four groups: sham, model (ischemia/reperfusion, I/R, MI group), I/R combined with ST36 (ST36 group), and I/R combined with sham acupuncture (MI+sham group). The MIRI model was established by ligation and reperfusion of the left anterior descending coronary artery. EA stimulation was applied at Neiguan (PC6) and Zusanli (ST36) in the ST36 group. Echocardiography, Evans blue-TTC double staining, ELISA and multi-omics techniques were adopted to evaluate therapeutic efficacy and analyze molecular characteristics. The results demonstrated that EA markedly reduced myocardial infarct size, improved cardiac systolic function, and decreased serum levels of IL-6, IL-1, and TNF-α as well as the expression of high mobility group box 1 (HMGB1) in the myocardial tissue. A total of 6 676 proteins were identified via differential proteomics analysis. One hundred and eighty-seven differentially expressed proteins were identified between the MI group and the sham group, among which the expression of 141 proteins was reversed after EA intervention. These proteins were mainly enriched in signaling pathways related to mitochondrial autophagy and apoptosis. Metabolomics analysis screened out 454 differential metabolites, which were predominantly involved in sphingolipid metabolism and alanine-aspartate-glutamate metabolism pathways. This study indicates that EA alleviates myocardial damage in the rat model of MIRI by suppressing inflammatory responses and regulating signaling pathways associated with myocardial structural maintenance, energy metabolism, and cell survival. The therapeutic mechanism of EA involves the synergistic modulation of multiple targets and pathways, providing experimental evidence for the comprehensive treatment of MIRI. 本研究旨在探究电针干预心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury, MIRI)的保护效应及其分子机制。选取40只SD雄性大鼠，随机分为4组，分别为空白组(sham)、I/R模型组(MI)、I/R+电针组(ST36)和I/R+假针组(MI+sham)。采用冠状动脉左前降支结扎后再开通的方法构建MIRI模型，EA组针对内关穴和足三里穴实施电针并给予电刺激;运用超声心动图、伊文思蓝(Evans blue)-红四氮唑(2,3,5-triphenyltetrazolium chloride, TTC)双染、ELISA以及多组学技术评估治疗效果并分析分子特征。结果显示电针能够显著减小大鼠心肌梗死面积，提升心脏收缩功能，降低血清中IL-6、IL-1、TNF-α水平以及心肌组织中高迁移率族蛋白1 (high mobility group protein b1, HMGB1)的表达;差异蛋白质组学检测共鉴定出6 676种蛋白质，MI组与Sham组之间存在187个差异蛋白，经电针干预后，141个蛋白的表达得到逆转，这些蛋白主要富集于线粒体自噬和凋亡等信号通路;代谢组学分析筛选出454个差异代谢物，主要集中于鞘脂代谢、丙氨酸-天冬氨酸-谷氨酸代谢等代谢通路。本研究提示电针通过抑制炎症反应，调控与心肌细胞结构维持、能量代谢及细胞存活相关的信号通路，进而改善MIRI大鼠的心肌损伤，作用机制涉及多靶点、多通路的协同调控，为MIRI的综合治疗提供了实验依据。.