Transcriptomic and Network Pharmacology Analysis of Targets Associated with Osteoporosis Improvement by Longzhong Sunshang Capsule and Experimental Validation.

INTRODUCTION: Osteoporosis (OP) severely impacts morbidity, mortality, and quality of life in affected individuals. This study aimed to screen key targets of Longzhong Sunshang Capsule for the treatment of OP via network pharmacology. METHODS: Transcriptomic and target data were retrieved from public databases, followed by differential expression analysis, machine learning, expression validation, and functional analyses (immune microenvironment, enrichment, molecular docking, molecular dynamics, toxicology, ELISA). RESULTS: This study identified FOXO3, XDH, and PLG as key OP targets (validated by ROC curves, AUC > 0.7), primarily enriched in chromatin remodeling and ribosome pathways. XDH interacted with the largest number of active components and showed strong binding affinity with luteolin (-10.0 kcal/mol). Key active components had liver damage toxicity scores of 0.62- 0.72. ELISA detection revealed that serum concentrations of FOXO3, XDH, and Plg were all significantly higher in osteoporosis patients than those in healthy controls (p < 0.0001). DISCUSSION: This study identified FOXO3, XDH, and PLG as key OP targets, highly expressed and diagnostically valuable, correlated with immune molecules, and enriched in chromatin remodeling and ribosomes. XDH has high diagnostic potential. Limitations include lack of in vitro/in vivo validation and insufficient FOXO3 mechanism characterization. CONCLUSION: FOXO3, XDH, and PLG are promising diagnostic targets, revealing the molecular mechanism underlying the anti-OP efficacy of Longzhong Sunshang Capsule.