Yishen Qianggu Mixture formulation suppresses osteometabolic inflammation by modulating the IL-17/NF-κB/MAPK signaling axis

ETHNOPHARMACOLOGICAL RELEVANCE: Yishen Qianggu Mixture (YSQGM) is a clinical empirical formula derived from Traditional Chinese Medicine (TCM) theory of "benefiting the kidney and strengthening bones," traditionally used to treat postmenopausal osteoporosis (PMOP) associated with kidney yang deficiency. Preliminary studies have shown that YSQGM improves bone mineral density (BMD), yet its integration of traditional efficacy with modern anti-inflammatory and bone metabolism-regulating mechanisms remains unclear. AIM OF THE STUDY: To elucidate YSQGM's anti-inflammatory effects and mechanisms on PMOP, with emphasis on verifying its inflammation-regulating role via multi-omics analysis and experimental validation. METHODS: An ovariectomy (OVX)-induced PMOP model was established in 36 Sprague-Dawley (SD) rats randomly assigned to six groups. Rats received YSQGM or alendronate by gavage for 8 weeks. BMD and trabecular microarchitecture were assessed by micro-computed tomography (Micro-CT), while serum bone turnover markers and inflammatory cytokines were measured by ELISA. Untargeted metabolomics and network pharmacology were integrated to identify differential metabolites and construct protein-protein interaction networks. Key targets were further examined by molecular docking, molecular dynamics (MD) simulations, and in vivo/in vitro validation. RESULTS: Medium and high doses of YSQGM significantly increased BMD, concomitantly decreased markers of bone formation and bone resorption, and ameliorated high-turnover PMOP. Untargeted metabolomics identified 46 differential metabolites and pinpointed ISL as a key anti-inflammatory constituent, with enriched pathways including triglyceride metabolism and the pentose phosphate pathway. Molecular docking and MD simulations predicted high affinity of ISL for PTGS2, MAPK14, GSK3B, and c-Fos. In vivo and in vitro experiments confirmed that YSQGM dose-dependently downregulated the expression of inflammation-related proteins (PTGS2, MAPK14, GSK3B, c-Fos, JNK, ERK, and NF-κB p65) and reduced osteoclast activity. CONCLUSION: YSQGM ameliorates bone metabolic imbalance in PMOP by suppressing proinflammatory mediators in the IL-17/NF-κB/MAPK signaling axis, providing novel mechanistic evidence for the use of TCM in PMOP prevention and treatment.