Integrated Chemical Profiling, Network Pharmacology, and Zebrafish/Cellular Models Identify Potential Anti-Osteoporotic Constituents of Cynomorium songaricum Rupr

Cynomorium songaricum Rupr. (CSR) is extensively utilized in traditional Chinese medicine for the treatment of bone disorders; however, its bioactive components and underlying mechanisms remain inadequately characterized. This study investigated the potential anti-osteoporotic constituents and mechanisms of CSR by integrating UPLC-Q-TOF-MS/MS, zebrafish and MC3T3-E1 cellular models, network pharmacology, molecular dynamics (MD) simulations, and RT-qPCR validation. Phenotypic evaluations showed that CSR aqueous extract significantly mitigated vertebral bone mineralization loss in glucocorticoid-induced zebrafish, while promoting cell proliferation and ALP activity in dexamethasone-treated MC3T3-E1 osteoblasts. Chemical profiling identified 21 components in the extract. Five candidate active ingredients and key targets were screened via network pharmacology, with predicted enrichment in the HIF-1 and VEGF signaling pathways. Molecular docking and 100 ns MD simulations predicted favorable binding poses and conformational stability between candidate compounds and core targets. Furthermore, maslinic acid, a candidate bioactive triterpenoid, improved cellular readouts under glucocorticoid challenge and was associated with reduced Casp3 mRNA and increased Runx2, Vegfa, and Mapk3 mRNA. These findings provide valuable insights into the multi-component osteoprotective potential of CSR, providing a solid scientific foundation for its future mechanistic exploration and therapeutic development.