Key Toxic Genes and Mechanisms of Di(2-ethylhexyl) Phthalate-Induced Non-Alcoholic Fatty Liver Disease: A Multi-Omics Study

INTRODUCTION: Non-alcoholic fatty liver disease (NAFLD) has emerged as the most prevalent chronic liver disease, with di(2-ethylhexyl) phthalate (DEHP) potentially exacerbating its progression. The toxicological mechanisms through which DEHP induces NAFLD remain elusive. This study aims to identify critical toxic genes involved in the development of DEHP-induced NAFLD. METHODS: The carcinogenic potential of DEHP was predicted based on its Simplified Molecular Input Line Entry System (SMILES) notation. Biomarkers were identified through differential expression analysis, and a nomogram was constructed and validated. The functional roles of these biomarkers were explored through enrichment analysis, immune cell infiltration, molecular docking, and molecular dynamics simulations. Single- cell analysis pinpointed the key cellular players in the process. RESULTS: The study identified activating transcription factor 3 (ATF3) and mitogen-activated protein kinase 8 (MAP3K8) as critical biomarkers. Pathways enriched by these biomarkers included "electron transfer from variant/mutation-inactivated PINK1 to complex I". Both ATF3 and MAP3K8 showed significant negative correlations with M2 macrophages and strong positive associations with activated mast cells. Binding affinities of ATF3 and MAP3K8 to DEHP were calculated at -5.9 kcal/mol and -7.7 kcal/mol. Hepatic stellate cells were ultimately identified as pivotal in the disease mechanism. DISCUSSION: Compared to traditional research approaches, this study employs network toxicology and molecular docking techniques, integrating single-cell sequencing and spatial transcriptomics to provide a comprehensive framework for investigating the potential toxic effects of di(2-ethylhexyl) phthalate (DEHP) in non-alcoholic fatty liver disease (NAFLD). CONCLUSION: This study provides valuable insights into the molecular processes underlying DEHP-induced NAFLD.