Department of Occupational and Environmental Health, College of Public Health, Xuzhou Medical University.
Environ Health Prev Med. 2024;29:46. doi: 10.1265/ehpm.24-00079.
Atrazine (ATR), a commonly used herbicide, is linked to dopaminergic neurotoxicity, which may cause symptoms resembling Parkinson's disease (PD). This study aims to reveal the molecular regulatory networks responsible for ATR exposure and its effects on dopaminergic neurotoxicity based on an integration strategy.
Our approach involved network toxicology, construction of protein-protein interaction (PPI) networks, gene ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, as well as molecular docking techniques. Subsequently, we validated the predicted results in PC12 cells in vitro.
An integrated analysis strategy indicating that 5 hub targets, including mitogen-activated protein kinase 3 (Mapk3), catalase (Cat), heme oxygenase 1 (Hmox1), tumor protein p53 (Tp53), and prostaglandin-endoperoxide synthase 2 (Ptgs2), may play a crucial role in ATR-induced dopaminergic injury. Molecular docking indicated that the 5 hub targets exhibited certain binding activity with ATR. Cell counting kit-8 (CCK8) results illustrated a dose-response relationship in PC12 cells. Real-time quantitative polymerase chain reaction (RT-qPCR) displayed notable changes in the expression of hub targets mRNA levels, with the exception of Mapk3. Western blotting results suggested that ATR treatment in PC12 cells resulted in an upregulation of the Cat, Hmox1, and p-Mapk3 protein expression levels while causing a downregulation in Tp53, Ptgs2, and Mapk3.
Our findings indicated that 5 hub targets identified could play a vital role in ATR-induced dopaminergic neurotoxicity in PC12 cells. These results provide preliminary support for further investigation into the molecular mechanism of ATR-induced toxicity.
莠去津（ATR）是一种常用的除草剂，与多巴胺能神经毒性有关，可能导致类似于帕金森病（PD）的症状。本研究旨在基于整合策略揭示ATR 暴露及其对多巴胺能神经毒性影响的分子调控网络。
我们的方法涉及网络毒理学、蛋白质-蛋白质相互作用（PPI）网络构建、基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析以及分子对接技术。随后，我们在体外 PC12 细胞中验证了预测结果。
综合分析策略表明，5 个关键靶标，包括丝裂原活化蛋白激酶 3（Mapk3）、过氧化氢酶（Cat）、血红素加氧酶 1（Hmox1）、肿瘤蛋白 p53（Tp53）和前列腺素内过氧化物合酶 2（Ptgs2），可能在 ATR 诱导的多巴胺能损伤中发挥关键作用。分子对接表明 5 个关键靶标与 ATR 具有一定的结合活性。细胞计数试剂盒-8（CCK8）结果表明，PC12 细胞中存在剂量反应关系。实时定量聚合酶链反应（RT-qPCR）显示关键靶标 mRNA 水平表达发生显著变化，除 Mapk3 外。Western blot 结果表明，ATR 处理 PC12 细胞导致 Cat、Hmox1 和 p-Mapk3 蛋白表达水平上调，而 Tp53、Ptgs2 和 Mapk3 蛋白表达水平下调。
我们的研究结果表明，鉴定的 5 个关键靶标可能在 ATR 诱导的 PC12 细胞多巴胺能神经毒性中发挥重要作用。这些结果为进一步研究 ATR 诱导毒性的分子机制提供了初步支持。
