CYP3A triggers BDE47-induced ferritinophagy and ferroptosis in spermatogenic cells through ROS-mediated m⁶A regulation of ATG12

Cytochrome P450 CYP3A (CYP3A) is among the most abundant hepatic cytochrome P450 subfamilies and also mediates the metabolism and toxicity of xenobiotics. Previous studies have reported that CYP3A is also expressed in the testis; however, its role and molecular mechanism in mediating male reproductive damage remain unclear. In this study, through in vitro and in vivo experiments, we demonstrated the role of CYP3A in 2,2',4,4'-tetrabromodiphenyl ether (BDE47)-induced reproductive toxicity. The results showed that BDE47 induced CYP3A expression in mouse testes, leading to oxidative stress and ferroptosis through excessive reactive oxygen species (ROS) and ferrous iron (Fe²⁺) overload, which was demonstrated by CYP3A overexpression or knockdown experiments in GC-2 cells. Mechanistically, in addition to direct ROS generation during metabolic processing, ferritinophagy contributed to intracellular Fe²⁺ accumulation. Specifically, BDE47-induced ROS was associated with reduced N6-methyladenosine (m⁶A) modification of Atg12 mRNA and increased autophagy-related (ATG12) expression, thereby promoting ferritin heavy chain 1 (FTH1) degradation and subsequent Fe²⁺ overload. These results were further validated by experiments using hydrogen peroxide or antioxidants in GC-2 cells, as well as by Atg12 haploinsufficiency in mice. Our findings demonstrate that CYP3A plays a critical role in male reproductive toxicity induced by BDE47.