Mechanism of action of HuPoSan on EMs rats as revealed by multi-omics combined with network pharmacology

BACKGROUND: Endometriosis (EMs) is a persistent inflammatory condition whose three main symptoms, mass, pain, and infertility, severely affect female patients of childbearing age. Hu-Po-San (HPS), a preparation consisting of various herbal medicines, is clinically used to treat EMs with positive results; however, the underlying mechanism of HPS's efficacy in EMs is still unclear. OBJECTIVES: This study employed network pharmacology, transcriptomics, non-targeted metabolomics, and experimental validation to elucidate the mechanism of HPS in the treatment of EMs. METHODS: A rat model of EMs was established to evaluate the therapeutic efficacy of HPS. Network pharmacology integrated with multi-omics was used to predict potential mechanisms, which were verified in vivo. Molecular docking was performed to explore interactions between major HPS constituents and core pathway proteins. RESULTS: HPS decreased ectopic lesion weight and volume in rats with EMs. Network pharmacology integrated with multi-omics analyses consistently suggested the Hippo signaling pathway as a key mechanism; concordantly, HPS activated this pathway in vivo and reversed disease-associated abnormalities-limiting proliferation and angiogenesis, suppressing inflammatory mediators, and restoring apoptosis-thereby inhibiting lesion progression. CONCLUSION: HPS limits ectopic lesion growth in EMs rats, likely via modulation centered on Hippo signaling pathway. The current work offers fresh perspectives on the methodical application of traditional Chinese medicine (TCM) to the treatment of EMs in rats. It provides a basis for developing new therapeutic approaches for treating EMs with HPS and its active ingredients.