Epidemiological studies have reported a progressive decline in parameters of male reproductive health. Increasing evidence has linked this decline to environmental contaminants. Per- and polyfluoroalkyl substances (PFAS), a major class of synthetic organic chemicals containing fully or partially fluorinated alkyl chains, are one of important environmental contaminants. In China, hexafluoropropylene oxide dimer acid (GenX) and chlorinated polyfluoroalkyl ether sulfonic acids (F-53B) are predominant PFAS. Reproductive toxicity of PFAS has been well-supported by evidence. However, evidence on male reproductive toxicity induced by GenX and F-53B remains limited. The present study aimed to explore the potential molecular targets and mechanisms of male reproductive toxicity induced by GenX and F-53B. Utilizing network toxicology, the intersection of targets between GenX and F-53B and male reproductive toxicity was identified. Their Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathways were further analyzed. Next, four core genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), tumor necrosis factor (TNF), MYC proto-oncogene (MYC), and estrogen receptor 1 (ESR1), were selected, and their relationships with the primary enrichment results were examined. Molecular docking and binding energy analysis were used to support strong interactions between the compounds and the core genes. The involvement of the core genes in toxic effects caused by GenX and F-53B was further supported by in vivo evidence. Overall, an integrated framework combining network toxicology, molecular docking, and in vivo validation was used to demonstrate male reproductive toxicity associated with GenX and F-53B. Mechanistic hypotheses were generated that may inform future investigations and clinical risk mitigation strategies.

Keywords: F-53B; GenX; male reproductive toxicity; molecular docking; network toxicology