Integrated network toxicology and molecular docking are used to elucidate the mechanism by which chlorpyrifos induces hepatotoxicity (Cyprinus carpio)

• CPF hepatotoxicity was enriched in TP53, ferroptosis, lipid and glutathione process. • TP53/SLC7A11/GPX4 signaling contributed to CPF-induced hepatocyte death. • Se could reverse CPF- and Erastin-induced ferroptosis and mitochondrial dysfunction. • GSH/GPX4 axis was the critical node in Se resisting CPF-induced lipid peroxidation. Chlorpyrifos (CPF), a typical organophosphorus pesticide, is frequently applied for pest control in households and agriculture worldwide. Liver, being the primary site for xenobiotic metabolism, is extremely vulnerable to drug threats. To elucidate the hepatotoxic mechanism of CPF and identify targets for its relief, the overlapping data between CPF-associated targets and liver injury-related genes were analyzed. It was found that CPF hepatotoxicity targets were mainly involved in oxidoreductase activity, fatty acid metabolism, glutathione metabolic process, response to biotic stimulus, ferroptosis, and mitochondrion disorders. TP53 molecular exhibited the strong binding stability with CPF, and TP53-mediated SLC7A11/GPX4 signaling suppression contributed to CPF-induced hepatotoxicity in Cyprinus carpio . CPF disrupted iron homeostasis by downregulating FTH, NCOA4, and FPN expressions, while simultaneously increasing ACSL4 expression to promote the accumulation of lipid peroxides MDA and LPO. Meanwhile, CPF reduced intracellular GPX4 levels, thereby diminishing lipid ROS scavenging capacity and triggering ferroptosis. Selenium (Se) could reverse CPF- and Erastin-induced ferroptosis and mitochondrial dysfunction. GSH depletion and GPX4 silencing reversed the alleviating effect of Se on CPF-induced lipid peroxidation, but they had little effect on iron contents. Meanwhile, Se addition also increased GSH contents and GPX4 protein expressions. The results revealed some targets of CPF-induced hepatotoxicity. These studies provided the theoretical basis for the future development of environmental monitoring and pollutant control strategies.