Network Pharmacology and In Vivo Validation Reveal Therapeutic Effects of Sutaehwan in Polycystic Ovary Syndrome by Modulating AMH-AMHR2 Signaling Pathway

This study aimed to investigate the therapeutic effects and underlying mechanisms of Sutaehwan (STH), a traditional herbal formula, in polycystic ovary syndrome (PCOS), with a focus on anti-Müllerian hormone (AMH)-driven ovarian dysfunction. A network pharmacology approach was used to predict STH-related molecular targets and their intersection with PCOS-associated genes. GO and KEGG enrichment analyses were conducted on 45 overlapping genes. Then, in vivo validation was performed in a letrozole-induced PCOS rat model treated with various doses of STH (0.45, 0.9, 1.8 mg/kg) or metformin (500 mg/kg). Key endpoints included estrous cycle, ovarian morphology, serum hormone levels, and expression of AMH/AMHR2. Network analysis revealed significant enrichment in pathways related to ovarian steroidogenesis and cell cycle regulation. Daily administration of letrozole induced classical PCOS phenotypes including increased cystic follicles, elevated AMH/testosterone levels, and disrupted estrous cycles. STH treatment dose-dependently restored ovulatory function, reduced cystic structures, and normalized corpus luteum volume and endometrial thickness. STH also significantly downregulated AMH and AMHR2 expression at both of transcriptional and protein levels, particularly at the 1.8 mg/kg dose, with effects comparable to those of metformin. STH exerts therapeutic effects in a PCOS model by targeting AMH-mediated ovarian dysfunction. Through combined systems pharmacology and experimental validation, this study supports the potential of STH as a multi-target, endocrine-modulating therapy for PCOS, particularly in cases characterized by elevated AMH activity.