Exploring the Key Targets and Underlying Mechanisms of Brain Injury Induced by the Plasticizer DEHP through Network Toxicology and Molecular Docking Approaches

Di(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer with documented neurotoxic effects, posing significant risks to brain tissue development and function. However, its precise molecular mechanisms and toxicological networks remain incompletely understood. This study aimed to elucidate the molecular mechanisms underlying DEHP-induced brain damage through network toxicology and molecular docking approaches. Public databases, including ChEMBL, STITCH, GeneCards, TTD, and OMIM, were used to construct the intersection network of DEHP-related targets and brain injury targets. Key pathways were identified via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Core targets were further screened using STRING and Cytoscape tools. To evaluate the diagnostic value of these core targets in brain injury, a logistic regression-based diagnostic model was developed, and a nomogram was constructed. The diagnostic performance and predictive accuracy of the nomogram were assessed using calibration curves, receiver operating characteristic (ROC) curves, and decision curve analysis (DCA). Immune cell infiltration in brain-injured and non-brain-injured cohorts was analyzed using the CIBERSORT method, and the correlation between core genes and immune cells was explored. Molecular docking techniques were employed to validate the binding affinities of DEHP to the core targets. The findings revealed KRAS, BCL2, EGFR, CCND1, CASP3, IL6, HSP90AA1, and ESR1 as pivotal targets of DEHP-induced brain injury. Enrichment analyses suggested that DEHP aggravates brain damage by disrupting cell survival, inflammation, apoptosis, and oxygen homeostasis. Key implicated pathways included the PI3K-Akt, JAK-STAT, FoxO, TNF, and HIF-1 signaling pathways. The nomogram model incorporating these eight core targets demonstrated high accuracy, as evidenced by calibration and ROC curve analyses. Immune infiltration studies revealed significant correlations between core targets and immune cell populations. Molecular docking confirmed that DEHP efficiently binds to core targets such as KRAS, BCL2, and IL6, supporting the hypothesis that it mediates brain injury via key signaling pathways. This study provides a comprehensive understanding of DEHP's neurotoxic mechanisms, offering valuable insights into its molecular basis and contributing to the future development of toxicological research and environmental risk assessment frameworks.邻苯二甲酸二(2-乙基己基)酯（Di(2-ethylhexyl) phthalate，DEHP）是一种应用广泛的塑化剂，已被证实具有神经毒性，对脑组织发育与功能构成显著风险。然而，其确切的分子机制与毒理网络仍未完全阐明。本研究旨在通过网络毒理学与分子对接技术，阐明DEHP诱导脑损伤的分子机制。研究借助ChEMBL、STITCH、GeneCards、TTD及OMIM等公共数据库，构建DEHP相关靶点与脑损伤靶点的交集网络；通过基因本体论（Gene Ontology，GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）富集分析筛选关键通路；利用STRING与Cytoscape工具进一步筛选核心靶点。为评估核心靶点在脑损伤中的诊断价值，本研究构建了基于逻辑回归的诊断模型，并绘制列线图（nomogram）；通过校准曲线、受试者工作特征（Receiver Operating Characteristic，ROC）曲线及决策曲线分析（Decision Curve Analysis，DCA）评估列线图的诊断性能与预测精度。采用CIBERSORT方法分析脑损伤与非脑损伤队列的免疫细胞浸润情况，并探索核心基因与免疫细胞的相关性。运用分子对接技术验证DEHP与核心靶点的结合亲和力。研究结果显示，KRAS、BCL2、EGFR、CCND1、CASP3、IL6、HSP90AA1及ESR1为DEHP诱导脑损伤的关键靶点。富集分析表明，DEHP可通过破坏细胞存活、炎症反应、细胞凋亡及氧稳态加重脑损伤，涉及的关键通路包括PI3K-Akt、JAK-STAT、FoxO、TNF及HIF-1信号通路。纳入上述8个核心靶点的列线图模型经校准曲线与ROC曲线分析证实具有较高诊断准确度。免疫浸润研究揭示核心靶点与免疫细胞群体间存在显著相关性。分子对接实验证实DEHP可与KRAS、BCL2及IL6等核心靶点高效结合，印证了其通过关键信号通路介导脑损伤的假说。本研究全面阐明了DEHP的神经毒性机制，为其分子基础研究提供了重要见解，同时为毒理学研究与环境风险评估框架的未来发展提供了参考依据。