Effect of hesperidin on anti-inflammation and cellular antioxidant capacity in hydrogen peroxide-stimulated human articular chondrocytes

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 μM) 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 μM) has a deleterious effect on normal cells, whilst low Hsd (5 and 10 μM) 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-1β, TNF-α, 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.