Integrated network toxicology and transcriptomics analysis reveals the pathogenic mechanisms in DBP/MBP-induced MASLD/MASH.

Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) and Metabolic Dysfunction-Associated Steatohepatitis (MASH) are challenging public health worldwide, with multifactorial pathogenic mechanisms. This study systematically employed network toxicology, molecular docking, molecular dynamics simulations and transcriptomics to investigate the metabolic toxicity mechanisms of the environmental pollutant dibutyl phthalate (DBP) its metabolite monobutyl phthalate (MBP). Protein-protein interaction network analysis identified 7 core proteins associated with MASLD and 5 with MASH, with ALBUMIN, PPARγ, HSP90AA1, and PTGS2 emerging as central hubs in DBP/MBP-induced hepatic metabolic disruption. Molecular docking experiments revealed strong DBP/MBP-key targets binding affinity, while molecular dynamics simulations demonstrated stable binding. RNA-seq profiling of DBP/MBP-exposed hepatocytes revealed significant transcriptomic alterations. KEGG pathway and GSEA analysis predictions with RNA-seq data pinpointed primary pathways potentially influenced by DBP/MBP, highlighting its multi-faceted impact on hepatic function. Furthermore, in vitro experiments validated the interaction of DBP/MBP with critical targets and demonstrated their ability to disrupt hepatocyte lipid homeostasis and promote lipid accumulation. This study provides mechanistic evidence linking DBP/MBP exposure to MASLD/MASH progression and lays a foundation for developing targeted therapeutic approaches.