Identifying the Chemical Profile and Pharmacological Mechanism of Xinbao Pill against Myocardial Ischemia-Reperfusion Injury Using Ultra-Fast Liquid Chromatography-Quadrupole-Time-Of-Flight Tandem Mass Spectrometry Coupled with Network Pharmacology-Based

Abstract Background: Xinbao Pill (Xin-Bao-Wan, XBW) is a traditional Chinese herbal formula, which is widely used in clinical treatment for cardiovascular diseases; however, the therapeutic effect of XBW on myocardial ischemia-reperfusion injury (MI/RI) is unclear. In this study, we evaluated the cardioprotective effect and molecular mechanism of XBW against MI/RI.Methods: A phytochemistry-based network pharmacology analysis was used to uncover the mechanism of XBW against MI/RI. Firstly, ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) method was used to identify chemicals in XBW. MI/RI-related targets of XBW were predicted using TargetNet database, OMIC database and reported literatures. GO and KEGG pathway enrichment analyses were performed using String database. Left anterior descending artery (LAD) ligation-induced MI/RI rat model and oxygen glucose deprivation-reperfusion (OGD/R)-induced H9c2 cell model were used to evaluate the cardioprotective effect and potential mechanism of XBW.Results: 37 chemicals were identified in XBW by using UHPLC-QTOF-MS; 50 MI/RI-related targets of XBW were predicted using TargetNet database, OMIC database and reported literatures. In vivo study, XBW (180 mg/kg) significantly reduced myocardial infarct size and creatine kinase MB (CK-MB) level induced by MI/RI in rat model; in vitro study, XBW protected H9c2 cells against OGD/R injury in a dose-dependent manner. GO and KEGG pathway enrichment analyses showed that the cardioprotective effect of XBW was associated with 5 significant pathways including autophagy, apoptosis, HIF-1 signaling pathway, PI3K-Akt signaling pathway, FoxO signaling pathway. Experimental investigation also verified that XBW could suppress apoptosis, autophagy and endoplasmic reticulum (ER) stress.Conclusion: XBW showed therapeutic effects against MI/RI mainly via attenuating apoptosis though suppressing excessive autophagy and ER stress.