Piperlongumine (PLM) is a natural product from the pods of a plant call Piper longum and was reported to possess antibacterial, antiangiogenic, neuroprotective, anti-inflammatory and anti-tumor activities. However, the role of PLM in inflammatory responses in human Osteoarthritis (OA) chondrocytes has not been yet explored. Thus, in this study, we investigated the anti-inflammatory action of PLM in human OA chondrocytes. Our results demonstrated that PLM treatment effectively reversed IL-1β-inhibited cell viability in a dose-dependent manner. In addition, PLM significantly inhibited the production of NO and PGE2, iNOS and COX-2 expression, as well as suppressed the production of MMP-3 and MMP-13 in IL-1β-stimulated human OA chondrocytes. Furthermore, PLM significantly prevented IL-1β-induced NF-κB activation in human OA chondrocytes. In conclusion, these findings demonstrated that PLM attenuated inflammatory responses in human OA chondrocytes stimulated by IL-1β, possibly through the NF-κB signaling pathway. Thus, PLM may serve as a potential anti-inflammatory agent in the treatment of OA.