啶虫脒对多能干细胞成神经分化的干扰作用  

Effects of Acetamiprid on Neuronal Differentiation of Pluripotent Stem Cells

作　　者：陈晓峰 朱宇澄 李明翰 郎玥明 郭俊岩 王晨曦 刘薇[1] CHEN Xiaofeng;ZHU Yucheng;LI Minghan;LANG Yueming;GUO Junyan;WANG Chenxi;LIU Wei(Key Laboratory of Industrial Ecology and Environmental Engineering,School of Environment,Dalian University of Technology,Dalian 116024,China)

机构地区：[1]大连理工大学环境学院,工业生态与环境工程教育部重点实验室,大连116024

出　　处：《生态毒理学报》2025年第2期17-24,共8页Asian Journal of Ecotoxicology

基　　金：国家重点研发计划项目(2023YFC39005201);国家自然科学基金(22376020);第17届Mandom国际动物实验替代研究基金。

摘　　要：尽管新烟碱类杀虫剂具有种属特异性毒性,但流行病学研究表明暴露于啶虫脒(acetamiprid,ACE)导致儿童神经发育受损,动物实验表明产前暴露于ACE导致子代小鼠运动功能异常。目前对于新烟碱类杀虫剂对哺乳动物的神经毒性和机制知之甚少。本研究基于多能干细胞体外神经分化模型,研究了0.5~50μmol·L^(-1)的ACE暴露对骨髓间充质干细胞的神经毒性效应潜能及毒性机制。首先对ACE与可能的作用靶点进行分子对接模拟,结果表明ACE与3种人烟碱型乙酰胆碱受体(nAChRs)以及人PPARγ受体均有一定的结合亲和性,虽然相互作用能低于其与昆虫AChBP受体的相互作用能,但可能通过作用于多个受体影响人体健康。在未造成显著细胞毒性浓度下,ACE暴露显著抑制鼠间充质干细胞(mesenchymal stem cells,MSCs)成神经分化的突触生成,ACE经鼠肝微粒体酶进行体外模拟代谢后该效应无显著变化。免疫荧光染色结果显示,50μmol·L^(-1)的ACE可显著抑制鼠神经元标志蛋白微管相关蛋白2(microtubule-associated protein 2,MAP2)的表达。0.5~50μmol·L^(-1)的ACE均可以显著提高细胞内活性氧水平。进一步用ACE作用于人源MSCs,对突触生成产生抑制作用,且在0.5~50μmol·L^(-1)暴露浓度范围内均刺激细胞钙瞬变,并呈剂量依赖性,诱导钙瞬变峰值比在1.25倍~1.5倍之间。本研究从细胞分化层面为流行病学发现和哺乳动物毒理学发现提供了机制解释,提示ACE可能干扰多能干细胞的谱系定向和命运决定。氧化应激及钙稳态失衡可能是ACE造成神经毒性的重要机制。须进一步筛查ACE损伤神经系统的敏感靶标,为该类农用化学品的风险管控提供科学依据。Despite the species-specific toxicity of neonicotinoid insecticides,epidemiological studies have indicated that exposure to acetamiprid(ACE)results in neurodevelopmental impairments in children,while animal experiments have demonstrated that prenatal exposure to ACE leads to motor function abnormalities in offspring mice.The neurotoxicity and mechanisms of neonicotinoid insecticides in mammals remain poorly understood.In this study,we investigated the potential neurotoxic effects and toxicity mechanisms of ACE exposure at concentrations of 0.5-50μmol·L^(-1)on bone marrow mesenchymal stem cells,utilizing an in vitro neural differentiation model of pluripotent stem cells.First,molecular docking simulations of ACE with potential targets were conducted,revealing that ACE exhibits some binding affinity to three human nicotinic acetylcholine receptors(nAChRs)and the human PPARγreceptor.Although the interaction energy is lower than that with the insect AChBP receptor,it may impact human health by acting on multiple receptors.Exposure to ACE at concentrations that did not induce significant cytotoxicity markedly inhibited synaptogenesis during neuronal differentiation of rat mesenchymal stem cells(MSCs),and this effect was not significantly altered by in vitro mimetic metabolism of ACE by hepatic microsomal enzymes.Immunofluorescence staining indicated that 50μmol·L^(-1)ACE significantly reduced the expression of m icrotubule-associated protein 2(MAP2),a marker protein of neurons,while 0.5-50μmol·L^(-1)ACE significantly elevated intracellular reactive oxygen species levels.Furthermore,the inhibitory effect of ACE on synaptogenesis in human MSCs stimulated calcium transients in a dose-dependent manner within the range of 0.5-50μmol·L^(-1),with the peak ratios of induced calcium transients ranging from 1.25-to 1.5-fold.This study provides a mechanistic e xplanation for epidemiological findings and mammalian toxicological findings at the level of cellular differentiation,suggesting that ACE may interfere with linea

关 键 词：啶虫脒 多能干细胞 突触生成 氧化应激 钙瞬变 

分 类 号：X171.5[环境科学与工程—环境科学]