Integrative network toxicology and transcriptomic analysis reveals molecular mechanisms of zinc pyrithione induced sperm damage in zebrafish.

Zinc Pyrithione (ZPT), a broad-spectrum antimicrobial agent widely employed in marine antifouling coatings, industrial biocides, and consumer products, has raised growing concerns regarding its male reproductive toxicity. However, the exact mechanisms are still unclear. In this study, adult male zebrafish were kept for 14 days to adapt and subsequently exposed to 0.30 μM ZPT for 21 days. To elucidate the molecular basis of ZPT-induced sperm damage, network toxicology and transcriptomic profiling were integrated. Database mining (Gene Expression Omnibus, PubChem, PharmGKB) identified 483 overlapping targets linking ZPT exposure to sperm damage. Protein-protein interaction networks and functional enrichment analyses indicated critical involvement of apoptosis, oxidative stress response, TP53 signaling, and neuroactive ligand-receptor pathways. Transcriptomic validation in zebrafish further demonstrated dose-dependent dysregulation of key genes (tdo2b, ctsba, gdf2, slc45a2, ungb). Moreover, molecular docking revealed stable Z interactions between ZPT and proteins associated with these pathways. Collectively, these findings establish oxidative stress, apoptotic cascades, and endocrine disruption as central mechanisms underlying ZPT-induced sperm toxicity, thereby providing critical insights for environmental risk assessment and regulatory policy development.