Excessive levels of oxidative stress can induce a senescence-associated secretory phenotype in chondrocytes which is characterized by the secretions of inflammatory mediators. Citrus flavonoid hesperidin (Hsd) is known to have anti-inflammatory and antioxidant capacities. Accordingly, we demonstrated the antioxidant property of Hsd against hydrogen peroxide (H2O2)-induced oxidative stress damages to chondrocytes in this study. Primary human chondrocytes were cultured in media supplemented with Hsd (5, 10, 50 and 100 mu M) to evaluate the dose-dependent cytotoxicity. Subsequently, cells were stimulated by H2O2 and supplied with Hsd to study chondroprotective effects. Results showed that high-dose of Hsd (50 and 100 mu M) has a deleterious effect on normal cells, whilst low Hsd (5 and 10 mu M) supplements improved mitochondrial activity, cytotoxicity, proliferation kinetic, survival and cell senescence in H2O2-stimulated chondrocytes. Real-time PCR revealed that Hsd downregulated the mRNA levels of COX-2, IL-1 beta, TNF-alpha, MMP-3, MMP-9, and upregulated IL-10, TIMP-1, SOX9. Western blotting further revealed that Hsd modulates Foxo1, Foxo3 and Nrf2 signaling pathways. Furthermore, Hsd improved total antioxidant capacity, and restored superoxide dismutases and glutathione peroxidase activities in H2O2-stimulated chondrocytes. In conclusion, our findings suggest that Hsd has the potential to improve antioxidant capacity in human chondrocytes.