ScholarWorks@중앙대학교

초록

Triflumizole (TFZ) is a widely-used imidazole-based fungicide effective against a broad range of fungal infections. Although its obesogenic and endocrine-disrupting properties have been studied, the cytotoxic effects of TFZ on male reproductive cells and the underlying mechanisms remain poorly understood. This study demonstrates that TFZ inhibits GC-1 spermatogonia (spg) cell proliferation, a type B spermatogonia, with an IC50 of 30.5 μM after 24 h of exposure. TFZ markedly increases reactive oxygen species (ROS) accumulation, leading to apoptosis and autophagy at high concentrations. It promotes cell death by upregulating pro-apoptotic proteins, including Fas/CD95, and by inducing the cleavage of caspase-3, caspase-7 and poly (ADP-ribose) polymerase (PARP). Additionally, TFZ activates autophagy-related proteins such as ATG5, ATG7, and LC3A-II. Notably, co-treatment of N-acetyl-L-cysteine (NAC), a potent antioxidant, significantly mitigates TFZ-induced cytotoxicity by reducing ROS levels. This results in a substantial decrease in the activation of pro-apoptotic and pro-autophagic proteins, including Fas/CD95, cleavage of caspase-3 and caspase-7, PARP, and LC3A-II formation. Furthermore, NAC preserves GPR30 activity, leading to the recovery of PI3K (Tyr458) and AKT (Ser473) phosphorylation in response to TFZ-induced damage, thereby restoring cell survival and proliferation. These findings indicate that TFZ's strong male germ cell toxicity is mediated by excessive ROS generation, which can be alleviated by antioxidant such as NAC. This highlights the critical connection between ROS and male germ cell health and underscore the protective potential of antioxidant agents in counteracting TFZ-induced reproductive cytotoxicity. © 2024

키워드