Elucidating the metabolic mechanisms and active constituents of ZuoGui Wan in combatting postmenopausal osteoporosis: A metabolomics and network pharmacology approach

• ZGW improves bone density and microstructure in OVX rats, showing potential as a treatment for PMOP. • Using metabolomics and network pharmacology, 14 blood-entry components and 33 key metabolites were identified, highlighting the role of amino acid metabolism in ZGW's effects. • Fourteen serum components, including emodin and rutin, were linked to ZGW's bone metabolism benefits, helping reduce bone loss. • Network pharmacology identified 106 target genes for PMOP, with molecular docking showing strong binding between ZGW compounds and key bone remodeling targets. • Arginine biosynthesis, NOS, and glutamine pathways are crucial for ZGW's anti-osteoporotic effects, promoting bone regeneration. Postmenopausal osteoporosis (PMOP) is a systemic metabolic disease with an imbalance of bone resorption and bone formation coupling as the basic pathogenesis. ZuoGui Wan (ZGW) can effectively ameliorate bone loss associated with decreased estrogen levels, the metabolic mechanisms and active ingredients involved remain unclear. We evaluated the efficacy of ZGW on bone loss in ovariectomized (OVX) rats. The blood-entry components of ZGW were identified using ultra-high performance liquid chromatography-high resolution mass spectrometry. The metabolic mechanisms of ZGW against PMOP were elucidated through metabolomics. Components highly correlated with differential metabolites were identified via correlation analysis. The metabolic mechanism of ZGW against PMOP was further confirmed by combining network pharmacology and metabolomics results and constructing an in vitro cell model for validation. ZGW reversed OVX-induced bone loss by enhancing bone density and improving bone microstructural integrity. Initially, 14 prototype components were identified in the serum of OVX rats following ZGW intervention. Metabolomics analysis revealed 33 differential metabolites involved in pathways such as arginine biosynthesis, and the metabolism of arginine, proline, histidine, aspartate, and glutamate. Additionally, network pharmacology and molecular docking analyses further dissected the active ingredients of ZGW, constructing a "Compound-Targets-Metabolic pathway-Metabolites" network against PMOP. this study comprehensively revealed the metabolic mechanism and active ingredients of ZGW against PMOP for the first time, providing a new perspective on the study of the mechanism of ZGW against PMOP.