Mechanistic insights into the anti-osteoporotic effects of Tripterygium wilfordii via network pharmacology, molecular docking, and experimental validation

Tripterygium wilfordii (Lei Gong Teng), a traditional Chinese medicinal herb, has shown potential in promoting bone health, yet its anti-osteoporotic mechanisms remain poorly defined. This study explored the underlying mechanisms of Tripterygium wilfordii (TWH) through network pharmacology, molecular docking, and experimental validation. Active compounds were identified from the TCMSP database using oral bioavailability ≥ 30 % and drug-likeness > 0.18, while osteoporosis-related targets were collected from GeneCards, OMIM, and TTD. Key targets were screened through protein–protein interaction analysis, and molecular docking assessed binding affinities. In vitro assays using mouse bone marrow-derived macrophages evaluated cytocompatibility, osteoclast inhibition, and changes in gene and protein expression. Eighteen active compounds and 121 putative targets were identified, with kaempferol and triptolide emerging as core constituents. Thirty-seven overlapping targets were enriched in IL-17, TNF, and estrogen pathways, and PTGS2, STAT3, and ESR1 were identified as key nodes. Molecular docking revealed strong binding affinities between major compounds and core proteins. TWH extract inhibited osteoclast formation dose-dependently and significantly downregulated PTGS2 and p-STAT3 expression at mRNA and protein levels. These findings suggest that TWH exerts anti-osteoporotic effects by modulating inflammation-related and estrogen-mediated signaling pathways, supporting its potential as a multi-targeted therapeutic candidate for osteoporosis.