背景:阿特拉津(ATR),一种常用的除草剂,与多巴胺能神经毒性有关,这可能会导致类似帕金森病(PD)的症状。本研究旨在基于整合策略揭示ATR暴露的分子调节网络及其对多巴胺能神经毒性的影响。
方法:我们的方法涉及网络毒理学,蛋白质-蛋白质相互作用(PPI)网络的构建,基因本体论(GO),和京都基因和基因组百科全书(KEGG)途径分析,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。
结果:综合分析策略表明5个中心目标,包括丝裂原活化蛋白激酶3(Mapk3),过氧化氢酶(猫),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起关键作用。分子对接表明5个hub靶标表现出与ATR的一定结合活性。细胞计数试剂盒-8(CCK8)结果显示PC12细胞中的剂量-反应关系。实时定量聚合酶链反应(RT-qPCR)显示hub靶mRNA表达水平的显著变化,除了Mapk3。Western印迹结果表明,在PC12细胞中ATR处理导致Cat的上调,Hmox1和p-Mapk3蛋白表达水平,同时导致Tp53、Ptgs2和Mapk3下调。
结论:我们的研究结果表明,所鉴定的5个hub靶点可能在ATR诱导的PC12细胞多巴胺能神经毒性中起重要作用。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。
BACKGROUND: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.