Hesperidin increases antioxidant capacity and improves preantral follicle survival and development during in vitro culture of goat ovarian tissue

ABSTRACT The present study investigated the effects of hesperidin supplementation on follicular survival, activation, and growth, as well as on antioxidant capacity, hydrogen peroxide production, intracellular thiol levels, and HSP70 expression during the in vitro culture of caprine ovarian cortical tissue. Ovarian tissue fragments were either immediately fixed (uncultured control) or cultured for 1 or 7 days in α-MEM + alone (cultured control), α-MEM+ supplemented with 50 μg/mL ascorbic acid (positive antioxidant control), or α-MEM+ containing hesperidin at 12.5, 25, 50, or 100 μmol/L. Hesperidin exerted concentration-dependent effects, with the most favorable responses observed at 100 μmol/L. Specifically, hesperidin at 25, 50, and 100 μmol/L maintained follicular survival after 1 and 7 days compared with the uncultured control (P < 0.05), whereas 100 μmol/L also increased follicular survival relative to the cultured control after 7 days (P < 0.05). In addition, hesperidin at 50 and 100 μmol/L promoted greater follicular and oocyte growth than both the uncultured and cultured controls (P < 0.05). The highest concentration of hesperidin also increased the antioxidant capacity of the culture medium, elevated intracellular thiol levels, and enhanced HSP70 immunostaining compared with the cultured control (P < 0.05), indicating improved cellular protection against oxidative stress. Overall, these results suggest that hesperidin promotes the maintenance of redox homeostasis and activates cytoprotective mechanisms during the growth of follicles and oocytes. Therefore, hesperidin emerges as a promising strategy for optimizing culture systems in goats.