| Objective To investigate the underlying mechanism of plastrum testudinis (PT) in the treatment of osteoporotic fracture (OF) using network pharmacology and experimental validation. Methods The components of PT and their corresponding targets were obtained with the help of BATMAN database, and relevant targets of OF were obtained by searching Genecards and OMIM databases. Intersection targets were input into the STRING database to obtain protein-protein interaction (PPI) information. The constructions of PPI network and PT-compound-target-OF network were done with Cytoscape 3.7.2, and GO function enrichment and KEGG pathway enrichment analyses were finished with Cytoscape3.7.2 and R. Additionally, to verify the reliability of the network pharmacology findings, we also conducted in vivo experiments to vary the effect of PT on improving OF and in vitro experiments, which investigated the phenotypes of human peripheral blood monocytes (HPBMs) under the intervention of PT. Results Six active compounds and 342 targets of PT were obtained, including 34 targets correlated with OF. There were 802 results gained through GO function enrichment analysis (P<0.05), and 67 results through KEGG pathway enrichment analysis, including 22 relevant signaling pathways (P<0.05), which mainly involved in TNF, MARK, estrogen, NF-κB, and other signaling pathways. Notably, our in vivo experiments proved that PT could effectively treat the bone defects in OF model rats. Results of in vitro experiments showed that PT could inhibit RANKL-induced osteoclast differentiation in HPBMs. Conclusion By regulating inflammatory response, hormone metabolism, and cell cycle, PT may treat OF through numerous compounds, targets and pathways. The inhibition of osteoclast differentiation may play an important role in the mechanism of PT against OF. |