Exploring the Toxicological Impacts of DOTP Exposure on Pulmonary Arterial Hypertension via Network Toxicology, Virtual Knockout, Molecular Docking and Experimental Validation.

Environmental factors are significant contributors to the pathogenesis of pulmonary arterial hypertension (PAH). Di-(2-ethylhexyl) terephthalate (DOTP) is a plasticizer widely used in daily life and poses potential health hazards. Nevertheless, the molecular mechanisms underlying PAH development following long-term DOTP exposure remain poorly understood. The aim of this study was to investigate the molecular mechanisms linking DOTP exposure to PAH by integrating network toxicology, virtual knockout, molecular docking and experimental validation methods. We screened four core targets (AKT1, EGFR, KDR, SRC) of DOTP and performed scTenifoldKnk virtual knockout analysis. Functional enrichment indicated that AKT1, EGFR and KDR significantly perturbed pathways related to the regulation of pulmonary artery endothelial cell (PAEC) damage and migration. We subsequently used CCK8, LDH release, RT-qPCR and transwell assays to confirm that DOTP exposure can promote PAEC damage and migration, accompanied by the upregulation of AKT1, EGFR and KDR expression. Finally, molecular docking and molecular dynamics were performed to elucidate the interactions and conformational changes involving DOTP and target proteins. This study reveals the molecular mechanism underlying the occurrence and development of DOTP-induced PAH, providing a new theoretical framework for the investigation of the effects of environmental pollutants on PAEC dysfunction and the pathogenesis of PAH.