Modified Erxian Decoction ameliorates premature ovarian insufficiency via HIF-1α-mediated regulation of mitochondrial fission and oxidative stress.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional herbal medicines have long been used to regulate female reproductive function. Erxian Decoction, a classical formula in traditional Chinese medicine, has is widely used to treat ovarian dysfunction and menopausal disorders. The Modified Erxian Decoction (MED) is clinically used to improve ovarian function; however, the pharmacological basis and active component interactions underlying its therapeutic effects remain unclear. AIM OF THE STUDY: To investigate the protective effects of MED against premature ovarian insufficiency (POI) and elucidate its underlying pharmacological mechanisms and optimize an active compound combination derived from the formula. MATERIALS AND METHODS: A cyclophosphamide-induced POI mouse model and 4-hydroperoxycyclophosphamide (4-HC)-induced granulosa cell injury model were established. Ovarian morphology, follicular development, reproductive outcomes, endocrine function, oxidative stress markers, and mitochondrial function were evaluated. Network pharmacology, molecular docking, and molecular dynamics simulations were integrated with experimental validation to identify key bioactive components and targets. A uniform design was used to optimize a representative compound combination consisting of icariin, tanshinone IIA, and berberine (ITB). RESULTS: MED significantly improved ovarian morphology, preserved follicular architecture, and ameliorated endocrine dysfunction in mice with POI, with the medium dose showing the most prominent therapeutic effects. MED markedly reduced oxidative stress and inhibited excessive mitochondrial fission, accompanied downregulating of DRP1 and FIS1 expression. The optimized ITB combination exerted stronger cytoprotective effects against granulosa cell injury than individual compounds. Mechanistic studies indicated that HIF-1α functions as an upstream regulator coordinating antioxidant responses and mitochondrial homeostasis. CONCLUSION: MED alleviates POI through multicomponent and multi-target mechanisms by regulating oxidative stress, HIF-1α-associated signaling, and mitochondrial dynamics. Mechanistic evidence supporting the optimized ITB combination provides rationale for translating ethnopharmacological knowledge into modern multicomponent therapeutics.