Osteoarthritis (OA) is a common degenerative disease characterized by articular cartilage destruction, subchondral bone remodeling, ectopic osteophyte formation and synovitis. It is now recognized that the integrity of the underlying subchondral bone is crucial for the maintenance of the overlying articular cartilage. Therapeutic agents that can prevent subchondral bone loss are demonstrate potential in the prevention and treatment of OA. Diosmetin (DIOS; 3',5,7 -trihydroxy-4'-methoxy flavone), a natural flavonoid, has been shown to exert anti-oxidative, anti-inflammatory, anti-apoptotic and anticancer properties. In this study, we found that diosmetin suppressed the DMM-induced subchondral bone loss and reduced subsequent cartilage degradation in vivo. Cellular-based assays showed that diosmetin inhibited RANKL-induced osteoclast formation and bone resorption,but did not affect IL-1β-induced chondrocyte hypertrophy. Biochemical analyses demonstrated that the anti-osteoclastic effect of diosmetin was at least in part due to the suppression of RANKL-induced activation of the ERK, p38, and JNK MAPK signaling pathways. Collectively, our results show that diosmetin have potential as a therapeutic agent the treatment of abnormal subchondral bone loss and cartilage degradation associated with the onset of OA.