Abstract: Objective To investigate the protective effects of pretreatment with Mongolian medicine Xiaohuixiang Sanwei Tang (XHX) against myocardial ischemia-reperfusion injury (MIRI) in mice. Methods Seventy-two male C57BL/6J mice were evenly and randomly divided into six groups: a sham operation group (Sham), model group (Model), compound Danshen dripping Pill (CDDP, 0.11 g·kg^-1) positive control group, and low-dose (XHX-L, 0.65 g·kg^-1), medium-dose (XHX-M, 1.3 g·kg^-1), and high-dose (XHX-H, 2.6 g·kg^-1) XHX groups. An MIRI model was established by 30 minutes of ischemia followed by 24 hours of reperfusion. Cardiac function parameters were recorded, TTC staining was used to detect myocardial infarct size after 24 hours of MIRI, HE staining was performed to observe myocardial tissue structure and cell morphology, and Masson staining was conducted to evaluate myocardial fibrosis. Blood was taken from the heart before the serum was separated to detect the indexes of MIRI oxidative stress in each group. A hypoxia/reoxygenation (H/R) model of H9c2 cardiomyocytes was constructed, and the protective effects of XHX were confirmed at the cellular level by CCK-8 assay. Network pharmacology was used to predict the active components and potential targets of the drug, construct a protein-protein interaction (PPI) network, and perform GO functional annotation and KEGG pathway enrichment analysis on overlapping targets using databases. Myocardial tissue samples from the Sham, Model, and XHX-H groups were collected for transcriptome sequencing to identify and verify potential key targets. Results TTC staining showed that XHX reduced the infarct size in mice 24 hours after MIRI. ELISA and assay kit results suggested that XHX decreased the levels of creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), and malondialdehyde (MDA) while increasing superoxide dismutase (SOD) levels 24 hours after MIRI. HE staining revealed that XHX ameliorated myocardial pathological changes following MIRI. Masson staining indicated that XHX effectively reduced the degree of myocardial fibrosis in mice with MIRI. H9c2 cell experiments confirmed that interventions with the XHX-M group effectively alleviated H/R-induced cardiomyocyte injury and exerted a significant protective effect against cell viability. Transcriptome sequencing combined with network pharmacology analysis revealed that XHX could regulate the progression of myocardial ischemic disease through chemokines. Conclusion XHX pretreatment alleviates MIRI-induced damage, reduces myocardial infarct size and fibrosis, and protects the heart in mice following MIRI.

Key words: Mongolian Medicine, Xiaohuixiang Sanwei Tang, Myocardial Ischemia-Reperfusion, Myocardial Infarction, Fibrosis, Inflammatory Response, Oxidative Stress, Mice