基于血清代谢组学和网络药理学探究复方丹参软胶囊改善心肌缺血小鼠的作用及机制OA

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.