Wubie Fanchun Formula-inducible metabolites in primary ovarian insufficiency model mice that facilitate ovarian renovation.

Abstract

CONTEXT: Premature ovarian insufficiency (POI), defined as ovarian activity cessation before age 40, significantly impacts both fertility and long-term health, with currently inadequate treatment options available. Wubie Fanchun Formula (WBFC) has been clinically used for over 20 years, showing benefits in TCM symptoms and hormone levels in POI patients with liver-kidney deficiency. However, its active components and mechanistic basis are still largely unknown. OBJECTIVE: To characterize the active ingredients and delineate the pharmacological basis of WBFC's action in managing POI. MATERIALS AND METHODS: LC-TOF-MS was used to identify WBFC's chemical constituents. Network pharmacology analysis pinpointed candidate mechanisms, which were tested in a 4-VCD-induced POI mouse model. Metabolic dysfunction was characterizedpyruvate tolerance tests, liver glycogen staining, and untargeted metabolomics. Key metabolites regulated by WBFC in model mice, phosphatidylinositol 38:5 (PI) and glutamine (GLN), were supplemented to validate their roles in POI pathology. RESULTS: 25 bioactive constituents in WBFC were found by LC-TOF-MS analysis. WBFC ameliorated ovarian dysfunction and activated the PI3K/AKT/FOXO3a pathway in parallel with simultaneously improving systemic metabolic parameters. Metabolomics identified PI and GLN among the top 30 most significantly altered metabolites, both showing marked depletion in POI mice and substantial restoration following WBFC treatment. Functional validation through exogenous supplementation demonstrated that combined PI and GLN administration effectively rescued ovarian dysfunction and specifically reactivated the ovarian PI3K-AKT pathway in 4-VCD induced POI mice. DISCUSSION AND CONCLUSION: Amino acids, saponins, anthraquinones, and carbohydrates are the main chemical components of WBFC. WBFC ameliorated POI and activated PI3K/AKT/FOXO3a signaling through metabolic regulation, with PI and GLN identified as critical therapeutic mediators.