Integrative network pharmacology and experimental validation to decipher the anti-angiogenic mechanisms of Xuandan Sanjie decotion in adenomyosis

OBJECTIVE: To investigate the mechanisms by which Xuandan Sanjie decoction (, XSD) inhibits angiogenesis in adenomyosis (AM), using both AM-derived primary endometrial stromal cells (ESCs) and a mouse model. METHODS: Core components and targets of XSD for AM treatment were predicted using multiple databases. Gene ontology and kyoto encyclopedia of genes and genomes pathway analyses were used to elucidate the potential mechanisms. Ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was applied to identify chemical constituents of XSD. A cobalt (II) chloride (CoCl2)-induced hypoxia-mimicking ESC model and an AM mouse model were established to experimentally validate the anti-angiogenic effects of XSD. RESULTS: According to bioinformatics analysis, hypoxia signaling and angiogenesis are crucial for AM progression. Twenty chemical constituents of the 106 compounds identified from the Traditional Chinese Medicine systems pharmacology database and analysis platform database were validated by UHPLC-MS/MS. Network pharmacology identified protein kinase B alpha, epidermal growth factor receptor, hypoxia inducible factor-1α (HIF-1α), and estrogen receptor 1 (ESR1) as core targets. According to in vitro and in vivo experiments, XSD exhibits anti-angiogenic properties through ESR1/G protein-coupled estrogen receptor (GPER)-mediated HIF-1α signaling. CONCLUSION: Our study confirms that XSD inhibits angiogenesis in AM viaESR1/GPER-mediated HIF-1α signaling. These findings elucidate the molecular mechanisms underlying XSD's therapeutic effects for abnormal uterine bleeding associated with adenomyosis, offering new therapeutic strategies for this condition.