F-53B exposure induced testicular premature aging through ZBP1-mediated programmed necrosis

Chlorinated polyfluorinated ether sulfonic acid (F-53B), a widely adopted substitute for perfluorooctane sulfonate (PFOS), which is one of the per- and polyfluoroalkyl substances (PFAS) chemicals, has garnered significant scientific attention due to its environmental persistence, bioaccumulation, and multi-organ toxic effects. As a central organ of the male reproductive system, the testis is essential for maintaining reproductive function. However, the mechanistic basis of F-53B-induced testicular toxicity remains largely uncharacterized. Therefore, we employed both in vitro (GC-1 spermatogonia cells and TM4 Sertoli cells) and in vivo murine exposure experiments to investigate the testicular toxicity of F-53B. Our results demonstrated that F-53B exposure induced cellular senescence and inflammatory damage in testicular cells, accompanied by marked increases in reactive oxygen species (ROS) accumulation. Mechanistic investigations revealed that F-53B triggered mitochondrial dysfunction and abnormal accumulation of Z-DNA, thereby initiating the ZBP1/RIPK3/MLKL-mediated programmed necrosis pathway, which ultimately promoted cellular senescence. In vivo experiments found that F-53B induced testicular inflammation and senescence. Collectively, this work reveals that F-53B induced testicular aging and provides an important basis for the evaluation of reproductive toxicity of F-53B.