Abstract: Objective To explore the mechanism of Achranthis Bioentatae Radix (AB) and Angelicae Pubescentis Radix (AP) in the treatment of osteoarthritis (OA).Methods In this study, the OA-related targets were screened out through network pharmacology. Then, a traditional Chinese medicine-active ingredient-protein network, a protein-protein interaction network and a core gene network were established. The resultant core genes were applied for KEGG function analysis. Furthermore, an interleukin (IL)-1β-induced model of OA cells was constructed. Cell viability was detected by CCK-8 assay. The expression of signaling proteins and gens were detected by Western blot and RT-PCR.Results A total of 14 active compounds, 35 potential OA targets, and 15 core genes were obtained from the drug pair of AB-AP. Among them, vascular endothelial growth factor A (VEGFA) and tumor protein 53 (TP53) were important nodes in the network and pathways of AB-AP, and might also be a key pathway for AB-AP to treat OA. Furthermore, the transcriptome data analysis of human OA samples showed that VEGFA and TP53 significantly decreased in the cartilage of OA patients, and the two genes were significantly correlated. The IL-1β-induced OA cell model showed that AB-AP significantly activated the VEGFA-TP53 pathway, while significantly reducing the gene and protein expression of MMP13 and up-regulating collagen Ⅱ gene and protein expression.Conclusions AB-AP may mainly up-regulate the VEGFA-TP53 pathway,in order to affect other multiple pathways, reduce the key proteins in the development of OA, and exert its anti-OA effect.