Hesperidin attenuates cyclophosphamide-induced urinary bladder injury: Sex-dependent histopathological and cellular alterations in rats.

Background

AND

Objective

Cyclophosphamide (CP) is a widely used chemotherapeutic agent whose clinical utility is limited by dose-dependent urotoxicity. This study investigated the sex-specific protective effects of hesperidin (HSP) against CP-induced bladder injury in rats, integrating biochemical, histological, and immunohistochemical outcomes. METHODOLOGY Adult male and female rats were allocated to control, HSP, CP, and HSP + CP groups (n = 8 per group). Oxidative stress and antioxidant status were assessed by malondialdehyde (MDA), reduced glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD). Bladder architecture and fibrosis were evaluated using hematoxylin and eosin and Mallory's trichrome staining, while epithelial integrity and proliferative activity were examined by E-cadherin and Ki-67 immunoexpression, respectively. Data were analyzed using a two-way ANOVA to assess the effects of treatment and sex, as well as their interaction.

Results

CP induced marked urotoxicity in both sexes, as evidenced by elevated MDA levels, depletion of GSH, CAT, and SOD, urothelial denudation, inflammation, collagen deposition, reduced E-cadherin expression, and increased Ki-67 immunoexpression. Female rats were more susceptible to CP-induced bladder injury, exhibiting greater mucosal damage, fibrosis, proliferative activity, collagen accumulation, higher MDA levels, and stronger Ki-67 expression than males. Hesperidin co-administration markedly attenuated CP-induced oxidative stress in both sexes. In male rats, hesperidin decreased MDA levels by 35.8% while increasing GSH, CAT, and SOD activities by 129.3%, 105.2%, and 18.5%, respectively, compared with the CP group. In female rats, the antioxidant effects were more pronounced, with MDA levels reduced by 73.9% and GSH, CAT, and SOD activities increased by 466.9%, 956.0%, and 203.2%, respectively, relative to the CP group. It also improved histoarchitectural alterations, reduced collagen accumulation, restored E-cadherin expression, and normalized Ki-67 levels toward control values, with a more pronounced protective effect in females. Hesperidin alone did not alter oxidative or structural parameters relative to controls.

Conclusion

Hesperidin co-administration substantially attenuated CP-induced bladder injury-reducing oxidative stress, improving histological architecture, limiting fibrosis, and restoring epithelial integrity and proliferative balance-with a more pronounced response in females; however, these benefits were partial, as most parameters did not fully return to control levels.