苯线磷和甲胺磷对TF-1细胞乙酰胆碱酯酶影响的差异  

Differential Effects of Fenamiphos and Methamidophos on Acetylcholinesterase in TF-1 Cells

作　　者：陈旸升 马永超 彭颖蓓 徐丽[1] 谢群慧[1,2] 赵斌 Chen Yangsheng;Ma Yongchao;Peng Yingbei;Xu Li;Xie Qunhui;Zhao Bin(State Key Laboratory of Environmental Chemistry and Ecotoxicology,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,Beijing 100085,China;College of Resources and Environment,University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院生态环境研究中心,环境化学与生态毒理学国家重点实验室,北京100085 [2]中国科学院大学资源与环境学院,北京100049

出　　处：《生态毒理学报》2023年第4期231-240,共10页Asian Journal of Ecotoxicology

基　　金：国家自然科学基金青年项目(22206202);国家自然科学基金重点项目(21836004);国家重点研发计划项目(2018YFA0901100)。

摘　　要：有机磷农药(organophosphorus pesticides,OPs)具有较强的神经毒性,主要是通过抑制胆碱能神经传导中的关键酶,乙酰胆碱酯酶(acetylcholinesterase,AChE,EC 3.1.1.7)的活性来实现的。苯线磷和甲胺磷是广泛用于农业生产的OPs,但对它们抑制人AChE活性的机制研究十分有限。本研究应用2种不同的给药方式,包括对培养的细胞和细胞裂解液进行药物处理,明确苯线磷和甲胺磷对人血液白血病细胞系TF-1中AChE酶活性的直接作用和对AChE基因转录表达的影响,从而揭示苯线磷和甲胺磷抑制AChE酶活性的机制与两者的差别。结果表明,高浓度苯线磷(10^(-3) mol·L^(-1))处理后,TF-1细胞活力降低,同时诱导细胞凋亡和坏死;而所有被试浓度的甲胺磷对细胞活力均没有明显影响。此外,对于TF-1细胞裂解液中的AChE,苯线磷和甲胺磷短期处理(1 h和5 h)均可产生直接抑制作用,其中苯线磷的抑制作用略强于甲胺磷(孵育1 h,IC_(50)值分别为1.181×10^(-6) mol·L^(-1)和2.837×10^(-6) mol·L^(-1))。对培养细胞的药物处理实验结果显示,苯线磷和甲胺磷(10^(-6) mol·L^(-1)和10-5 mol·L^(-1))处理24 h后,均显著降低了TF-1细胞的AChE酶活性,苯线磷的抑制率略高于甲胺磷。而与对酶活性的抑制作用相反,苯线磷和甲胺磷对TF-1中AChE H转录本表达有轻微的上调作用,以甲胺磷更为明显,提示存在反馈调节机制。总结上述结果,我们发现苯线磷对TF-1细胞中AChE的抑制作用总体略强于甲胺磷,而甲胺磷对AChE基因表达的反馈上调作用更明显。从而首次从对AChE酶的直接抑制作用和生物合成影响的不同角度阐述了2个OPs对AChE影响的差异,为进一步的分子机制研究提供了实验数据。Organophosphorus pesticides(OPs)have strong neurotoxicity,which is through the inhibition of acetylcholinesterase(AChE,EC 3.1.1.7),an enzyme critical for cholinergic neurotransmission.Fenamiphos and methamidophos are widely used OPs in agricultural production,but research on their mechanisms of inhibiting human AChE activity is limited.In this study,the effect of fenamiphos and methamidophos on AChE enzyme activity in human erythroleukemia cell line TF-1 and their impact on AChE gene transcriptional expression were clarified.The results showed that after treatment with high concentration of fenamiphos(10^(-3) mol·L^(-1)),the cell viability of TF-1 cells decreased,while inducing cell apoptosis and necrosis;however,all concentrations of methamidophos tested had no significant effect on cell viability.In addition,short-term(1 h and 5 h)treatment with fenamiphos and methamidophos lead to direct inhibitory effects on AChE in TF-1 cell lysate,with fenamiphos having a slightly stronger inhibitory effect than methamidophos(1 h incubation,IC_(50) value is 1.181×10^(-6) and 2.837×10^(-6) mol·L^(-1),respectively).The results of these two OPs on cultured cells showed that after 24 h of treatment with fenamiphos and methamidophos(10^(-6) mol·L^(-1) and 10-5 mol·L^(-1)),the AChE enzyme activity of TF-1 cells was significantly reduced,and the inhibition rate of fenamiphos was slightly higher than that of methamidophos.Contrary to the inhibitory effect on enzyme activity,both fenamiphos and methamidophos slightly upregulated the expression of AChE H transcripts in TF-1,with methamidophos being more obviously,indicating a feedback regulatory mechanism.Based on the above results,we found that the inhibitory effect of fenamiphos on AChE in TF-1 cells is generally slightly stronger than that of methamidophos,and there is feedback upregulation effect of methamidophos on AChE gene expression.Thus,for the first time,the differences in the effects of two OPs on AChE were elucidated from different perspectives of direct inhibition

关 键 词：苯线磷 甲胺磷 乙酰胆碱酯酶(AChE) 人血液白血病细胞TF-1 

分 类 号：X171.5[环境科学与工程—环境科学]