Osteoarthritis (OA) is the most common type of degenerative joint disease and secreted inflammatory molecules serve a pivotal role in it. Peimine has been reported to have anti-inflammatory activity. In order to investigate the potential therapeutic role of Peimine in OA, mouse articular chondrocytes were treated with IL-1 beta and different doses of Peimine in vitro. The data revealed that Peimine not only suppressed IL-1 beta-induced production of nitric oxide (NO) and prostaglandin E2, but also reduced the protein levels of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). In addition, Peimine inhibited the IL-1 beta-induced mRNA expression of matrix metalloproteinase (MMP)-1, MMP-3, MMP-9, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5. Furthermore, Peimine inhibited IL-1 beta-induced activation of the mitogen-activated protein kinase (MAPK) pathway. The protective effect of Peimine on IL-1 beta-treated chondrocytes was attenuated following activation of the MAPK pathway, as demonstrated by the increased expression levels of MMP-3, MMP-13, ADAMTS-5, iNOS and COX-2 compared with the Peimine group. The in vivo data suggested that Peimine limited the development of OA in the mouse model. In general, the data indicate that Peimine suppresses IL-1 beta-induced inflammation in mouse chondrocytes by inhibiting the MAPK pathway, suggesting a promising therapeutic role for Peimine in the treatment of OA.