Integrative network toxicology analysis reveals mechanistic links between PFOA exposure and male reproductive dysfunction

Abstract Objective To systematically investigate the molecular mechanisms linking perfluorooctanoic acid (PFOA) to male reproductive health outcomes within a network toxicology framework, and to compare shared pathways across erectile dysfunction (ED), male infertility, and testicular dysfunction. Methods PFOA associated targets were identified using CTD, SwissTargetPrediction, and TargetNet, while disease related genes were collected from GeneCards. Protein–protein interaction (PPI) networks were constructed using STRING and analyzed in Cytoscape to identify hub genes. GO and KEGG enrichment analyses were performed to characterize biological processes and pathways affected by PFOA. Molecular docking was used to assess binding affinities between PFOA and key target proteins. In addition, preliminary in vitro experiments were performed to examine the expression patterns of selected candidate genes following PFOA exposure. Results A total of 105 PFOA related genes were identified. Intersection analyses yielded 15, 25, and 42 genes associated with ED, male infertility, and testicular dysfunction, respectively. Ten core genes (AR, ESR1, BCL2, NOS1, NOS2, NOS3, ACE, SIRT1, TLR9, SHBG) were shared across all phenotypes and showed high connectivity in the PPI network. Enrichment analyses indicated involvement in reproductive system development, hormone and nuclear receptor regulation, calcium and nitric oxide (NO) signaling, mitochondrial and redox processes, and apoptosis and inflammation related pathways. Preliminary in vitro experiments further validated the expression changes of selected key genes. Conclusions This study identifies key molecular targets and pathways potentially mediating PFOA associated male reproductive toxicity. The results support a working hypothesis that PFOA associated male reproductive toxicity may involve altered androgen estrogen receptor signaling, NO related pathways, and apoptosis/inflammation associated regulation. These findings provide a theoretical basis for future mechanistic validation and risk assessment.