OBJECTIVE To investigate the in vivo active components of Xinkeshu tablets against myocardial ischemia.

METHODS Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was employed to analyze the chemical profile of the Xinkeshu tablets methanol extract and rat serum containing the drug. Network pharmacology was utilized to predict the potential mechanisms of the serum-absorbed components against myocardial ischemia. Screening for active components was performed using cellular and zebrafish models.

RESULTS A total of 114 compounds, including flavonoids, phenolic acids, terpenoids, and tanshinones, were identified in the methanol extract of Xinkeshu tablets. Analysis of drug-containing rat serum led to the identification of 116 compounds, comprising 37 parent compounds and 79 metabolites. Network pharmacology results indicated that the primary serum-absorbed components of Xinkeshu tablets may exert anti-myocardial ischemia effects by modulating key signaling pathways such as PI3K-AKT, AGE-RAGE, MAPK, and Ras, via targeting critical proteins including SRC, PIK3R1, PIK3CA, HSP90AA1, MAPK1, STAT3, HRAS, and AKT1. Compounds such as caffeic acid, dihydrocaffeic acid, protocatechuic acid, Danshensu, and daidzein significantly restored cell viability, alleviated cytotoxicity, increased mitochondrial membrane potential, and mitigated pericardial edema and bradycardia in zebrafish.

CONCLUSION This study preliminarily elucidated the main in vivo active components of Xinkeshu tablets against myocardial ischemia, providing an experimental basis for its clinical application and further development.