Investigation of ferroptotic cell death induced by acute alcohol exposure in rat testes: evaluation of the effects of ferrostatin and boric acid

Title: Investigation of ferroptotic cell death induced by acute alcohol exposure in rat testes: evaluation of the effects of ferrostatin and boric acid Aim: This study aimed to investigate whether ethanol exposure induces ferroptosis in testicular tissue and to evaluate the protective effects of boric acid and Ferrostatin-1 through the analysis of key ferroptosis-related markers. Method: A total of 48 male Sprague-Dawley rats, aged 3–4 months and weighing 200–250 g, were used in the present study. Ferroptotic processes in rat testicular tissues were evaluated following ethanol administration. The animals were randomly assigned into four experimental groups: ethanol only, ethanol + boric acid, ethanol + Ferrostatin-1, and ethanol + boric acid + Ferrostatin-1. Biochemical analyses were conducted to determine the levels of SLC7A11, GPX4, glutathione (GSH), and malondialdehyde (MDA). Histopathological evaluations were performed using Hematoxylin–Eosin (H&E) staining, while ultrastructural changes were examined by transmission electron microscopy (TEM). Results: Ethanol significantly suppressed SLC7A11 and GPX4 expression and reduced GSH levels while increasing MDA levels, indicating activation of ferroptosis. In contrast, both boric acid and Fer-1 reversed these effects by restoring GSH levels and reducing lipid peroxidation. Histological analyses confirmed severe testicular damage and ferroptotic features in the ethanol group, while reduced damage was observed in the boric acid and Fer-1 groups. However, combined treatment with both agents resulted in unexpected adverse effects. Conclusion: Ethanol activates ferroptosis in rat testicular tissue by disrupting antioxidant defenses and promoting lipid peroxidation. Boric acid and Ferrostatin-1 individually demonstrate therapeutic potential in mitigating ethanol-induced testicular ferroptosis, though their combined use may lead to undesirable interactions. Keywords: Alcohol, Testis, Ferroptosis, Boric acid, Ferrostatin-1