Riboflavin activates the Bmp4/Bmpr1α signaling to reverse arsenic-induced male reproductive toxicity: in vivo and in vitro studies

As a widely distributed hazardous substance in agriculture and food, arsenic has become a global public health concern. Riboflavin is known for its anti-inflammatory and antioxidant properties, but whether it mitigates arsenic-induced male reproductive damage remains unclear. To clarify arsenic’s reproductive toxicity, network toxicology was first used to identify potential targets. Subsequently, thirty-six 4-week-old male mice were administered arsenic trioxide (5 mg/L in drinking water) and/or riboflavin (10 mg/kg in diet) for 22 weeks to establish a riboflavin intervention model. Network toxicology identified that arsenic induces male infertility via multiple signaling pathways, such as the TGF-β signaling pathway. In vivo, arsenic diminished sperm quality, caused histopathological damage, and impaired both blood-testis barrier integrity and Sertoli cell proliferation, while riboflavin effectively mitigated these detrimental effects. In vitro, TM4 cell experiments further confirmed that riboflavin reversed arsenic-induced decline in cell viability and cytoskeletal damage. This study is the first to demonstrate that the protective effect of riboflavin is mainly attributed to its ability to mediate the binding of Bmp4 to Bmpr1α, thereby reversing arsenic-induced toxicity, providing a new perspective for exploring riboflavin’s role in alleviating arsenic toxicity.