Forchlorfenuron exposure Induces hepatocyte Apoptosis via MKK3/p38/ATF-2 signaling Pathway

Abstract Forchlorfenuron is a widely used plant cytokinin in traditional Chinese medicine and agricultural cultivation to boost resistance, postpone senescence, and increase productivity. However, improper forchlorfenuron use results in excessive residues and contamination, raising health and safety concerns. The in vitro toxicity of forchlorfenuron on HepaRG cells was investigated in our research. Results showed that forchlorfenuron inhibited HepaRG cell viabilities in a concentration and time-dependent manner. Forchlorfenuron induced cellular apoptosis and the increased intracellular reactive oxygen species (ROS) indicated the participation of oxidative stress. Molecular docking and network pharmacology data suggested that the hepatotoxicity of forchlorfenuron might involve the MAPK signaling pathway. After 24 hours of forchlorfenuron exposure, the p38-MAP kinase, upstream kinases MKK3, and the transcription factor ATF-2 was maximally activated. Apoptosis induced by forchlorfenuron was significantly reduced by pretreatment of the P38 inhibitor SB203580. These findings implicated that HepaRG hepatocyte injuries were generated by forchlorfenuron through the induction of cellular apoptosis via MKK3/p38/ATF-2 pathways. Forchlorfenuron application should be closely managed to prevent potential liver damage.