Systematic Investigation of the Potential Multi-Target Pharmacological Mechanisms of Astragaloside IV in Polycystic Ovary Syndrome via Network Pharmacology and In Vivo/In Vitro Experiments

Background Polycystic ovary syndrome (PCOS) is a prevalent endocrine-metabolic disorder affecting reproductive-aged women and is characterized by hyperandrogenemia, ovulatory dysfunction, and polycystic ovaries. Astragaloside IV (AS-IV), an active compound derived from Astragalus membranaceus, shows promise in the treatment of metabolic disorders. However, the precise molecular targets and mechanisms of action in PCOS remain unclear. This study aimed to elucidate the therapeutic effects and underlying mechanisms of AS-IV in PCOS. Methods AS-IV’s therapeutic effects of AS-IV were assessed in a rat model of PCOS. Potential AS-IV targets were predicted using the PharmMapper and SwissTargetPrediction databases and expanded using STRINGdb. PCOS-related differentially expressed genes (DEGs) were identified from GEO datasets, and weighted gene co-expression network analysis (WGCNA) revealed the disease-associated gene modules. Overlapping drug-disease targets were analyzed using protein-protein interaction (PPI) network, Gene Ontology (GO), and KEGG pathway enrichment. The core targets were validated using molecular docking. In vitro, qPCR was used to assess key gene expression in the control, DHT model, AS-IV, EP300 inhibitor (C646), and Nrf2 inhibitor (ML385) groups. Results A total of 371 potential AS-IV targets were identified. Analysis of GEO data yielded 2,286 DEGs, with WGCNA identifying key PCOS-related modules and hub genes. The intersection revealed 31 key targets, including five core genes. In vivo, AS-IV improved ovarian pathology, increased antioxidant enzyme levels, reduced inflammatory cytokine, testosterone, and LH levels, and increased estradiol levels. In vitro, the PCOS, EP300, and Nrf2 inhibitor groups showed decreased EP300, NFE2L2, HMOX1, and AKT1 expression and increased MMP9 expression compared to the controls and AS-IV group. Conclusion AS-IV ameliorated endocrine and ovarian abnormalities in PCOS by modulating the EP300/Nrf2/HMOX1/MMP9 axis, demonstrating multi-target antioxidant, anti-inflammatory, and hormone-regulatory effects, supporting its potential as a therapeutic agent for PCOS.