Ferulic acid relieves hypoxic damage by inhibiting endoplasmic reticulum stress, oxidative stress, and apoptosis through activating FOXA2-modulated Nrf2/HO-1 pathway in spermatogonial cells

Male infertility is a global public health problem affecting human development. Ferulic acid (FA) is an important active phenolic substance that has been found to exert a protective effect on male reproductive system. Here we aimed to investigate the effect of FA on hypoxia-induced spermatogenesis damage and explore the underlying mechanisms. Mouse spermatogenic GC-1 spg cells were cultured under hypoxic conditions to establish an in vitro model of spermatogenesis damage. The small interfering RNAs targeting Forkhead Box A2 (FOXA2) were transfected into GC-1 spg cells to construct FOXA2-silencing cells. Cell counting kit-8 (CCK-8) and flow cytometry assays were performed to detect cell viability and Apoptosis, respectively. Western blot analysis was conducted to examine the expression levels of related proteins. We found that FA improved the decreased cell viability of GC-1 spg cells under hypoxic conditions. FA attenuated hypoxia-induced endoplasmic reticulum (ER) stress, oxidative stress, and Apoptosis in GC-1 spg cells. Both mRNA and protein levels of FOXA2 were reduced after hypoxia induction, which were reversed by FA treatment. Knockdown of FOXA2 reversed the inhibitory effects of FA on ER stress, oxidative stress, and Apoptosis in hypoxia-exposed GC-1 spg cells. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) overexpression restored the role of FOXA2 knockdown in FA-treated GC-1 spg cells under hypoxic conditions. Overall, we demonstrated the inhibitory effect of FA on hypoxia-induced ER stress, oxidative stress, and Apoptosis in mouse spermatogenic GC-1 spg cells. Furthermore, FOXA2-mediated Nrf2 signaling contributed to the protective effects of FA against hypoxia-induced spermatogenesis damage. These findings suggest that FA may have the potential to ameliorate hypoxia-induced spermatogenesis injury.

FOXA2; Ferulic acid; Hypoxia; Male infertility; Spermatogenesis damage.