Integrative network toxicology and experimental validation reveal novel molecular mediators of omeprazole-induced nephrotoxicity

Omeprazole, a widely used proton pump inhibitor, has been increasingly linked to adverse renal outcomes such as acute kidney injury (AKI) and chronic kidney disease (CKD), yet its nephrotoxic mechanisms remain unclear. In this study, we utilized an integrative network toxicology approach, combined with molecular docking and in vitro validation, to elucidate potential mechanisms underlying omeprazole-induced renal toxicity. By intersecting omeprazole-related genes with those implicated in AKI and CKD, we identified 73 overlapping targets enriched in the PI3K-Akt signaling pathway, EGFR inhibitor resistance, and xenobiotic response. A protein–protein interaction network and topological analysis revealed six hub targets (CASP3, CCND1, EGFR, MMP9, PARP1, PPARG), all showing strong binding affinities to omeprazole in molecular docking simulations. Functional validation in HK-2 cells demonstrated that omeprazole reduced cell viability, induced morphological damage, and increased apoptosis in both acute (300 μM, 24 h) and chronic (20 μM, 7 days) exposure models. qPCR analysis confirmed significant upregulation of CASP3 , EGFR , MMP9 , PARP1 , and PPARG , with CCND1 exhibiting model-dependent expression changes. These findings provide new insights into the molecular basis of omeprazole-associated nephrotoxicity and suggest potential biomarkers and therapeutic targets for mitigating renal injury.