Mechanisms underlying the adverse health outcomes of dibutyl phthalate and bis(2-ethylhexyl) phthalate based on network toxicology, molecular docking, and molecular dynamics simulation

The widespread use of dibutyl phthalate (DBP) and di(2-ethylhexyl) phthalate (DEHP) has raised concerns about their potential toxicity to human health. This study aimed to identify the potential targets of DBP and DEHP and their related pathways. Potential target proteins were predicted by functional enrichment analysis and protein-protein interaction (PPI) networks were constructed to identify hub targets, while molecular docking and molecular dynamics (MD) simulations validated the interactions between DBP and their target proteins. The results showed that DBP/DEHP targets were significantly enriched in pathways, among which the lipid and atherosclerosis pathway was the primary focus of the present study, together with progesterone-mediated oocyte maturation, and prostate cancer. Four hub proteins associated with lipids and atherosclerosis, namely GSK3B, MAPK1, HSP90AA1, and SRC, were identified. Molecular docking results demonstrated strong interactions between DBP/DEHP and these four hub proteins. MD simulations further confirmed the effective binding of DBP to its two protein targets (GSK3B and HSP90AA1). By integrating network toxicology, molecular docking, and MD simulations, this study provides preliminary computational hypotheses and mechanistic clues for understanding the biological effects of DBP and DEHP. These in silico findings also offer theoretical support for prioritized pollutant risk assessment and toxic effect mechanism interpretation for phthalate contaminants.