目的 整合血清代谢组学和入血成分网络药理学探究复方丹参软胶囊（Compound Danshen Soft Capsule，CDSC）对异丙肾上腺素（isoprenaline，ISO）诱导的小鼠心肌缺血模型的改善作用及机制。方法 小鼠ip ISO建立心肌缺血模型，给予CDSC干预后，采用苏木素-伊红（hematoxylin-eosin，HE）和Masson染色评估心肌组织病理学变化；测定心肌组织能量代谢酶活力；ELISA法测定心肌组织中心肌损伤标志物水平；采用超高效液相色谱-飞行时间质谱（UHPLC-TOF-MS）对CDSC入血成分进行鉴定分析；使用CDSC差异入血成分进行网络药理学分析探究潜在作用机制；使用血清非靶向代谢组学分析CDSC对心肌缺血小鼠血清代谢水平的影响；采用Western blotting验证相关通路的蛋白表达。结果 CDSC显著减轻ISO诱导的心肌缺血小鼠心肌组织病理损伤，升高心肌组织中能量代谢酶活力（P＜0.001），降低心肌损伤标志物水平（P＜0.001）。共鉴定出17个差异入血成分，网络药理学分析筛选出187个与心肌损伤相关的交集靶点。京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析表明，CDSC改善心肌缺血的作用可能和表皮生长因子受体（epidermal growth factor receptor，EGFR）酪氨酸激酶抑制剂耐药性、腺苷酸活化蛋白激酶（AMP-activated protein kinase，AMPK）、晚期糖基化终产物及其受体（advanced glycation end products-receptor for advanced glycation end products，AGE-RAGE）等信号通路相关。血清代谢组学分析表明CDSC对心肌缺血小鼠的改善作用可能与胰高血糖素、碳代谢、丙酮酸代谢、嘌呤代谢等多个代谢途径有关。Western blotting结果进一步证实，CDSC可剂量相关性地调控AMPK、哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）蛋白的磷酸化水平（P＜0.01、0.001）。结论 CDSC通过调控能量代谢等方式改善心肌缺血，其作用机制可能与AMPK/mTOR信号通路有关。

Objective To investigate the ameliorative effect and underlying mechanism of Compound Danshen Soft Capsules (复方丹参软胶囊, CDSC) on isoproterenol (ISO)-induced myocardial ischemia in mice through the integration of serum metabolomics and network pharmacology of absorbed bioactive components. Methods A mouse model of myocardial ischemia was established by ip ISO. After CDSC intervention, the pathological changes in myocardial tissue were evaluated using hematoxylin-eosin (HE) and Masson staining. The activity of energy metabolism enzymes in myocardial tissue were measured. ELISA was used to measure the levels of myocardial injury markers in myocardial tissue. Identification and analysis of CDSC blood components were performed using ultra-high performance liquid chromatography-time-of-flight mass spectrometry (UHPLC-TOF-MS). CDSC differential blood components for network pharmacology analysis was used to explore potential mechanisms. Serum untargeted metabolomics was used to analyze the effect of CDSC on serum metabolic levels in mice with myocardial ischemia. Western blotting was used to validate the protein expressions of related pathways. Results CDSC significantly alleviated the pathological damage of myocardial tissue in ISO-induced myocardial ischemia mice, increased the activities of energy metabolism enzymes in myocardial tissue (P < 0.001), and reduced the levels of myocardial injury markers (P < 0.001). A total of 17 differential blood components were identified, and network pharmacology analysis screened 187 intersecting targets related to myocardial injury. Kyoto encyclopedia of genes and genomes enrichment analysis suggested that the effect of CDSCs in improving myocardial ischemia may be related to signaling pathways such as resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, AMP-activated protein kinase (AMPK) and advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE). Serum metabolomics analysis suggested that the improvement effect of CDSC on myocardial ischemia mice may be related to multiple metabolic pathways such as glucagon, carbon metabolism, pyruvate metabolism and purine metabolism. Western blotting results further confirmed that CDSC could dose-dependently regulate the phosphorylation levels of AMPK and mammalian target of rapamycin (mTOR) protein (P < 0.01, 0.001). Conclusion CDSC improves myocardial ischemia by regulating energy metabolism and other mechanisms, and its mechanism may be related to AMPK/mTOR signaling pathway.

R285.5

广东省科技计划项目（2024A0505090009）