Protective effects of nuclear factor erythroid 2-related factor on oxidative stress and apoptosis in the testis of mice before adulthood

Abstract Oxidative stress disrupts the intracellular redox balance that modulate many signaling pathways, including nuclear factor erythroid 2-related factor 2 (Nrf2)/Keap1 signaling. However, the antioxidant roles of Nrf2 in the testis before adulthood have not been reported. Accordingly, in this study, we aimed to investigate the effects of the Nrf2 antioxidant system on protection of testicular cells against oxidative stress at different stages of development in the testis of mice before adulthood. Male mice (1, 2, 4, and 8 weeks old) were used, and their relative testes weights were calculated. Malondialdehyde (MDA) contents and superoxide dismutase (SOD) activity were detected to evaluate the antioxidant capacity in the testes. Additionally, Nrf2 signaling pathway and mitochondrial apoptotic pathway proteins were evaluated by western blotting, and the localizations of Nrf2, protein gene product (PGP) 9.5, and activated-caspase 3 in testicular cells were examined using immunohistochemistry. The results showed that the activities of caspase-8 and caspase-3 and the number of activated-caspase 3-positive testicular cells per tubule were increased after 1 week of age. Moreover, MDA contents were increased and SOD activity was decreased with age in mouse testes before adulthood. The expression of PGP9.5 was increased, as well as the number of positive testicular cells per tubule. In addition, Nrf2 translocation to the nuclei of testicular cells also increased, accompanied by activation of the Nrf2/Keap1 signaling pathway. Moreover, nuclear factor-κB was inhibited, and the mitochondrial apoptotic pathway was activated in mouse testes before adulthood. Overall, our findings demonstrated that oxidative stress increased with age in mouse testes before adulthood and that oxidative stress could induce apoptosis in testicular cells. However, testicular cells are still in a rapid proliferative state owing to the antioxidant protection of Nrf2. Thus, our study provided new insights into oxidative stress-mediated impairment of spermatogenesis with age in mouse testes before adulthood and evidence for the protective role of Nrf2 in male fertility.