基于脂质代谢组学研究苏合香“温通心脉”改善心梗后线粒体能量代谢的作用机制

Storax has the effect of “warming heart meridian”. The related prescription has definite efficacy in the treatment of coronary heart disease and is widely used in clinical practice, but its myocardial protection mechanism is still not clear yet and there is a lack of in-depth research under the guidance of the holistic view. The previous results show that Storax can improve myocardial infarction injury and maintain mitochondrial morphology in rats with coronary heart disease. The preliminary results of metabolomics suggest that its role in myocardial protection is closely related to energy metabolism. Therefore, this project is to using metabonomics and molecular biology technique, under the guidance of the holistic view of traditional Chinese medicine, based on lipid biomarkers information and take AMPK/PGC1α pathway as the breakthrough point to explore network of “the efficacy - endogenous metabolites - molecular function pathway” and verify this hypothesis “Storax can exert myocardial protection by protecting mitochondrial structure and regulating lipid metabolism, thus improving mitochondrial energy metabolism disorder after myocardial infarction”, and then confirm that molecular mechanism of Storax on myocardial protection. It will provide scientific basis for its clinical application and optimization, and provide a reference for establishing mechanism research strategies of “warming heart meridian” of the traditional Chinese medicine against myocardial injury.

苏合香具有“温通心脉”之功效，相关方剂治疗冠心病临床疗效确切、应用广泛，但其心肌保护机制尚不清楚，缺乏整体观指导下深入研究。申请人前期研究证实苏合香可改善冠心病大鼠心肌梗死损伤，维持线粒体形态，代谢组学对内源性代谢物群进行初步解析，发现其药效干预靶点与能量代谢通路密切相关。申请人拟在中医整体观指导下，采用代谢组学结合分子生物学技术，以脂质生物标记物信息为基础，以AMPK/PGC1α信号通路为切入点，探索“药效-内源性代谢物群-分子功能途径”网络，验证“苏合香通过保护心梗后线粒体结构功能，调节脂质代谢，从而改善线粒体能量代谢紊乱发挥心肌保护作用”这一假说，继而确证苏合香心肌保护的分子机制，为其临床应用、优化升级提供实验依据，为中药“温通心脉”抗心肌损伤机制的研究策略提供借鉴。

目的：苏合香治疗冠心病临床疗效确切、应用广泛，但其心肌保护机制尚不清楚，缺乏整体观指导下深入研究。本研究从整体-细胞-分子水平阐明苏合香抗冠心病大鼠心肌梗死的作用机制，通过“药效-内源性代谢物群-分子功能途径”网络整体解析其是否通过改善线粒体能量代谢紊乱而发挥心肌保护作用，揭示AMPK/PGC1α通路在其的重要作用。方法：基于大鼠MI模型和心肌细胞I/H模型，采用ELISA、透射电镜、病理染色、代谢组学、免疫荧光、Western blot等实验方法验证其药效及作用机制。结果：（1）苏合香可改善大鼠心肌梗死面积与线粒体超微结构，减少心肌酶指标与细胞凋亡，从而改善能量代谢发挥心肌保护作用；（2）结合苏合香代谢网络靶点，应用多元统计学分析筛选出苏合香发挥心肌保护作用的内源性能量代谢物群，为苏合香心肌保护作用机制验证提供依据；（3）苏合香能够有效的改善p-AMPK，PGC1α，NRF1和TFAM这些与线粒能量代谢有关的蛋白的表达，加入AMPK抑制剂后，结果显示能逆转p-AMPK，PGC1α，NRF1和TFAM等蛋白的表达，说明苏合香可能是通过AMPK/PGC1α信号通路发挥作用；（4）网络药理学筛选预测冠心苏合丸心肌保护作用可能涉及的通路包括PI3K/Akt信号通路，HIF-1α信号通路，NF-kB信号通路等，并通过实验对PI3K/AKT通路进行验证，发现冠心苏合丸可促进PI3K-AKT磷酸化表达，从而抑制细胞凋亡发挥心肌保护作用。结论：本课题从整体-细胞-分子水平阐明苏合香治疗冠心病的作用机制，通过“药效-内源性代谢物群-分子功能途径”网络整体解析明确了苏合香通过改善线粒体能量代谢紊乱而发挥心肌保护作用，揭示AMPK/PGC1α通路在其的重要作用，为苏合香临床应用、优化升级提供实验依据，为中药“温通心脉”抗心肌损伤机制的研究策略提供借鉴。