Simplified assessment of acetylcholinesterase inhibition by environmental organophosphorus pesticides

IF 5.4 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Ruihong Zhu , Yiyun Liu , Yongchao Ma , Tong Xu , Guanglei Yang , Antonio Zandona , Tena Čadež , Zrinka Kovarik , Heidi Qunhui Xie , Li Xu , Bin Zhao
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Abstract

Environmental contaminants, such as pesticides, can inhibit the enzymatic activity of acetylcholinesterase (AChE), an enzyme necessary for neurotransmission. The inhibitory effects of structurally diverse pesticides on AChE may result from either reversible or covalent interactions. Therefore, assessing their potency typically requires different assay design to determine either dissociation constants or rate constants, respectively. To avoid complex kinetic experiments and enable comparison of potencies across structurally diverse pesticides, we optimized an AChE inhibitor detection system using an endpoint Ellman assay in 96-well plates. Given the significant interspecies variability in sensitivity to inhibitors, we investigated AChE inhibition using both electric eel AChE (eeAChE) and human AChE (hAChE). After confirming the repeatability, reproducibility, and solvent compatibility of the detection system using the reversible inhibitor BW284c51, we determined the inhibition potency of selected organophosphorus (OP) pesticides based on IC50 values. We found that chlorpyrifos, fenamiphos and ethoprophos were more potent inhibitors of hAChE than eeAChE. In contrast, phosalone and methamidophos showed similar inhibitory effects on both enzymes. The potencies aligned well with previously reported inhibitory rate constants. In conclusion, since OPs acts as progressive inhibitors of AChE, our optimized assay offers a simplified yet effective method for assessing their inhibitory potency. It also allows for comparative evaluation of various environmental pollutants based solely on IC50 values, eliminating the need for complex kinetic studies.
环境有机磷农药对乙酰胆碱酯酶抑制作用的简化评价。
环境污染物,如杀虫剂,可以抑制乙酰胆碱酯酶(AChE)的酶活性,乙酰胆碱酯酶是神经传递所必需的酶。不同结构的农药对乙酰胆碱酯酶的抑制作用可能是可逆的,也可能是共价的。因此,评估它们的效价通常需要不同的分析设计来分别确定解离常数或速率常数。为了避免复杂的动力学实验,并能够比较不同结构农药的效力,我们在96孔板上使用端点埃尔曼法优化了乙酰胆碱酯酶抑制剂检测系统。鉴于对抑制剂的敏感性存在显著的物种间差异,我们研究了电鳗AChE (eeAChE)和人类AChE (hAChE)对AChE的抑制作用。在确定了可逆抑制剂BW284c51检测系统的重复性、再现性和溶剂相容性后,我们根据IC50值确定了所选有机磷(OP)农药的抑制效力。我们发现毒死蜱、虫胺磷和乙氧丙磷对乙酰胆碱酯的抑制作用强于乙酰胆碱酯。相比之下,磷沙酮和甲胺磷对这两种酶的抑制作用相似。效价与先前报道的抑制速率常数一致。综上所述,由于OPs是乙酰胆碱酯酶的进行性抑制剂,我们优化的实验提供了一种简化而有效的方法来评估其抑制效力。它还允许仅基于IC50值对各种环境污染物进行比较评估,从而消除了复杂动力学研究的需要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
7.70
自引率
3.90%
发文量
410
审稿时长
36 days
期刊介绍: Chemico-Biological Interactions publishes research reports and review articles that examine the molecular, cellular, and/or biochemical basis of toxicologically relevant outcomes. Special emphasis is placed on toxicological mechanisms associated with interactions between chemicals and biological systems. Outcomes may include all traditional endpoints caused by synthetic or naturally occurring chemicals, both in vivo and in vitro. Endpoints of interest include, but are not limited to carcinogenesis, mutagenesis, respiratory toxicology, neurotoxicology, reproductive and developmental toxicology, and immunotoxicology.
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