Osteoarthritis (OA) is a common arthrosis characterized by degeneration and inflammation of articular cartilage. In recent decades, peiminine (Pm) has been identified as one of the active ingredients of Fritillaria plants. According to reports, Pm has a potent anti-inflammatory effect in various diseases. However, the effectiveness of Pm as an anti-inflammatory in OA has not previously been reported. This research aims to evaluate the anti-inflammatory effect of Pm on interleukin (IL)-1β-induced mice chondrocytes and its chondroprotective effect in a mouse OA model with surgical destabilization of the medial meniscus. IL-1β-induced expression of nitric oxide (NO), prostaglandin E2 (PGE2), cyclooxygenase-2 (COX-2), tumor necrosis factor alpha (TNF-α), inducible nitric oxide synthase (iNOS) and interleukin-6 (IL-6) were all inhibited significantly by Pm pretreatment in vitro. In addition, Pm also inhibited the expression of thrombospondin motifs 5 (ADAMTS-5) and matrix metalloproteinase-13 (MMP-13), which are responsible for the degradation of the extracellular matrix (ECM). Additionally, the degradation of aggrecan and collagen II was reversed by Pm. Furthermore, Pm inhibited Akt phosphorylation and the nuclear transfer of nuclear factor-κB (NF-κB) and activated Nrf2/HO-1 signaling pathways both in vitro and in vivo. These findings suggested that Pm alleviated inflammatory effects in the IL-1β-induced chondrocytes. Therefore, Pm might be a potential therapeutic agent for OA.