Bushen Huoxue Formula alleviates lipid accumulation in premature ovarian insufficiency by activating the LRP6/β-catenin signaling pathway.

Abstract

BACKGROUND: Bushen Huoxue Formula (BSHXF), a traditional Chinese medicine, effectively treats premature ovarian insufficiency (POI) by modulating of lipid metabolism. PURPOSE: This study aimed to investigate the underlying mechanism through which BSHXF improves POI by regulating lipid metabolism. METHODS: Network pharmacology identified potential pathways associated with BSHXF in POI. A POI rat model was induced using subcutaneous corticosterone injections and treated with BSHXF to assess its efficacy. The impact of BSHXF on lipid metabolism, autophagy, and the LRP6/β-catenin signaling pathway was evaluated in these rats. Additionally, palmitic acid (PA)-induced granulosa cells (KGN cells) were used in vitro. A LRP6 inhibitor (Salinomycin sodium salt, Sal) was applied to these cells, followed by a series of molecular biology experiments to elucidate the specific regulatory role of BSHXF on lipid metabolism. RESULTS: Network pharmacology analysis indicated that the Wnt/β-catenin signaling pathway is a potential target of BSHXF in POI. In vivo, BSHXF alleviated POI-like symptoms in rats, reduced lipid accumulation, suppressed excessive autophagy, and activated the LRP6/β-catenin signaling pathway. In vitro, Sal inhibited the ability of BSHXF-containing serum to activate LRP6/β-catenin signaling, reduce lipid accumulation, and counteract excessive autophagy in PA-induced KGN cells. This was accompanied by increased expression of LRP6 and β-catenin, enhanced lipogenesis, elevated lipid droplet formation, decreased lipid catabolism, and oxidation, and enhanced autophagic flux. Cycloheximide chase and Western blot analyses further confirmed that BSHXF stabilizes LRP6 by inhibiting its ubiquitin-proteasomal degradation. CONCLUSIONS: BSHXF activates LRP6/β-catenin signaling pathway by inhibiting the ubiquitination degradation of LRP6, thereby improving POI. This study reveals that BSHXF may improve POI by modulating LRP6 stability, thereby affecting lipid metabolism.