OBJECTIVE: To investigate molecular mechanisms of Jinwujiangu decoction (金乌健骨方, JWJG) in treating osteoarthritis (OA) using network pharmacology analysis. METHODS: Principal active compounds of JWJG were screened out via the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and BATMAN-TCM, and potential targets for OA treatment were identified through Online Mendelian Inheritance in Man (OMIM) and GeneCards databases. The JWJG network diagrams of both principal chemical compound-action targets and OA treatment target-OA disease were constructed applying the Cytoscape 3.7.2 software. The diagram of protein-protein interaction network was plotted for core analysis. Meanwhile, the common targets and relevant signaling pathways involved in both networks were analyzed using the Gene Ontology function analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment. The predicted results were ultimately verified through animal experiments. RESULTS: Effects of JWJG were indicated in acting on key targets interleukin-6, insulin, protein kinase B, glyceraldehyde-3-phosphate dehydrogenase, and mitosis-specific MRE11-RAD50-NBS1 associated protein by regulating signaling pathways of phosphoinositide 3-kinase- protein kinase B, mitogen-activated protein kinases, tumor necrosis factor, and colorectal cancer. Meanwhile, it inhibited the over-activation of signaling pathways and the release of inflammatory factors in OA treatment. Following a signaling pathway analysis utilizing network pharmacology technique, it was demonstrated that JWJG could treat OA through the Wnt/β-catenin signaling pathway verified by animal experiments. CONCLUSIONS: The present study preliminarily analyzed the pharmacological mechanism of JWJG in treating OA on a network pharmacology approach and provided a great theoretical significance for clinical application.