Traditionally, the development of osteoarthritis (OA) is associated with factors such as aging and injure, but more and more epidemiological and biological evidence suggests that the disease is closely related to metabolic syndrome and metabolic components. Ubiquitin-specific protease 3(USP3), a member of the USPs family, is a specific protease capable of cleavage of ubiquitin chains linked by proline residues. In our presented study, we firstly found that USP3 expression level was decreased in OA. USP3 over expression inhibited IL-1 beta induced chondrocytes apoptosis and nuclear factor kappa B (NF-kappa B) activation. USP3 knockdown induced chondrocytes apoptosis and activated NF-kappa B pathway. USP3 interacts with TRAF6 (tumor necrosis factor-receptor-associated factor 6), which is an essential adaptor protein for the NF-kappa B (nuclear factor kappa B) signaling pathway and plays important roles in inflammation and immune response. IL-1 beta treatment up-regulated the polyubiquitination of TRAF6 in chondrocytes, which was attenuated when USP3 was forced expression. Our study mechanistically links USP3 to TRAF6 in osteoarthritis development. Moreover, these data support the pursuit of USP3 and TRAF6 as potential targets for osteoarthritis therapies. (C) 2019 Published by Elsevier Inc.