4-tert-butylphenol triggers common carp hepatocytes ferroptosis via oxidative stress, iron overload, SLC7A11/GSH/GPX4 axis, and ATF4/HSPA5/GPX4 axis.

4-tert-butylphenol (4-tBP) is a toxic environmental pollutant with moderate bioaccumulation, environmental persistence, and long-term toxicity. Its toxicity to aquatic organisms has become an issue of concern. However, the molecular mechanism of 4-tBP toxicity to aquatic organisms remained unclear. Liver is a target organ for environmental pollutants. Here, we established 4-tBP-exposed toxicity model in vivo and primary hepatocyte model in vitro in common carp (Cyprinus carpio L.). We found increased hepatic-somatic index (HSI) and abnormal serum biochemical indexes (ALT, AST, and LDH) after 4-tBP exposure, indicating liver damage. We further revealed that 4-tBP damaged the structural integrity of the livers with typical features of ferroptosis. Based on toxicogenomics analysis, we found ferroptosis is likely to be involved in the mechanism of 4-tBP-induced liver damage. Moreover, our in vivo and in vitro experiment provided evidences that 4-tBP-exposure led to excess oxidative stress, iron overload, decreased MMP, and abnormal expression of ferroptosis-related factors. Interestingly, ferrostatin-1 (Fer-1, a ferroptosis inhibitor) pretreatment alleviated above changes. In summary, we demonstrated that 4-tBP triggered hepatocytes ferroptosis via oxidative stress, iron overload, SLC7A11/GSH/GPX4 axis, and ATF4/HSPA5/GPX4 axis. For the first time, we discovered that Fer-1 can ameliorate the toxicity of 4-tBP, which needs more investigations. Our results provided a scientific basis of molecular mechanism of 4-tBP-induced fish poisoning.