Abstract
OBJECTIVE: Dioctyl terephthalate (DOTP), a widely used plasticizer in food packaging and environmental materials, has raised concerns regarding its potential impact on human health. This study aims to investigate the neurotoxicity-related mechanisms of DOTP through an integrated approach combining network toxicology, molecular dynamics simulations, and in vivo validation. METHODS: We retrieved the DOTP chemical structure from PubChem and predicted potential protein targets using the Similarity Ensemble Approach (SEA), SwissTargetPrediction, and SuperPred. We performed protein-protein interaction (PPI) network analysis using STRING and Cytoscape to identify core neurotoxicity-associated targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted to characterize the biological relevance of the 90 intersecting targets. We used AutoDock to assess the binding affinity of DOTP to the core proteins, and employed GROMACS for molecular dynamics (MD) simulations to explore the stability and conformational dynamics of the docked complexes. In vivo experiments, encompassing behavioral assessments, histological examinations, and molecular assays, were conducted to evaluate the effects of DOTP on neurological function, neuronal integrity, and target pathway dysregulation. RESULTS: We identified 90 neurotoxicity-related targets, among which EGFR, BCL2, CASP3, MAPK8, TLR4, NFKB1, and MTOR emerged as core nodes within the PPI network. GO and KEGG analyses revealed the involvement of these targets in diverse biological processes, cellular components, molecular functions, and signaling pathways. Molecular docking indicated favorable binding affinities between DOTP and the identified core targets, a finding further supported by MD simulations. Moreover, DOTP-treated mice exhibited significant neurofunctional deficits and neuronal loss, accompanied by profound oxidative stress, neuroinflammation, and apoptotic activation, substantiating its potential neurotoxicity. CONCLUSION: Our findings provide a theoretical foundation for understanding the predicted molecular mechanisms of DOTP-induced neurotoxicity. The integration of computational modeling and in vivo phenotypic validation suggests that DOTP may pose neurological risks, highlighting the need for further experimental evaluation of plasticizer alternatives.