Integrated network toxicology and experimental validation identify candidate targets in phthalate-related metabolic-associated fatty liver disease.

Phthalate esters are pervasive endocrine-disrupting chemicals associated with metabolic disorders, but their contribution to metabolic-associated fatty liver disease (MAFLD) remains unclear. We investigated shared targets and pathways linking eight phthalate-related compounds to MAFLD using network toxicology, transcriptomic data mining, molecular docking, and cellular validation. Integrated analysis identified 75 predicted phthalate-related targets, including 41 MAFLD-associated genes enriched in lipid metabolism and inflammatory pathways. Cross-analysis of three GEO datasets prioritized 14 candidate genes, with PPARG, PTGS2, ESR1, PPARA, and BCL2 showing high network centrality. Molecular docking predicted favorable interactions between the tested compounds and candidate proteins, with monobenzyl phthalate (MBeP) showing the highest predicted affinity. In palmitic acid-oleic acid (PA-OA) -treated HepG2 cells, MBeP increased lipid accumulation, reactive oxygen species generation, apoptosis, and mitochondrial membrane depolarization and altered selected candidate genes. In primary mouse hepatocytes, MBeP similarly increased lipid accumulation and reactive oxygen species generation and altered PTGS2 and PPARA protein expression. These findings identify candidate targets and pathways potentially involved in MBeP-associated hepatic lipid dysregulation and support further mechanistic investigation of phthalate-related effects in MAFLD.