Mechanistic insights into chlorpyrifos-induced nephrotoxicity revealed by network toxicology and molecular docking

Chlorpyrifos (CPF), a widely used organophosphorus pesticide, poses potential nephrotoxicity risks, yet its molecular mechanisms remain incompletely elucidated. This study integrated network toxicology, molecular docking, molecular dynamics (MD) simulations, and in vitro experiments to systematically explore CPF-induced renal injury mechanisms. ADMET and toxicity predictions indicated CPF's potential nephrotoxic risk. Through cross-database screening, 33 overlapping targets were identified, and seven core targets, MPO, XDH, TLR4, NFE2L2, NOS2, NOS3, and SOD2, were further selected via protein-protein interaction network analysis. Enrichment analysis revealed significant involvement in oxidative stress, inflammatory response, and HIF-1 signaling pathways. Molecular docking and 100 ns MD simulations demonstrated stable binding of CPF and its metabolites to core targets. In vitro experiments using HK‑2 cells confirmed that CPF upregulated MPO expression and induced oxidative damage and inflammatory responses, which were attenuated upon MPO knockdown. This multi-level strategy reveals that CPF may trigger nephrotoxicity via oxidative stress and inflammatory activation, providing new insights into its toxicological mechanisms.