Ruihong Zhu , Yiyun Liu , Yongchao Ma , Tong Xu , Guanglei Yang , Antonio Zandona , Tena Čadež , Zrinka Kovarik , Heidi Qunhui Xie , Li Xu , Bin Zhao
{"title":"环境有机磷农药对乙酰胆碱酯酶抑制作用的简化评价。","authors":"Ruihong Zhu , Yiyun Liu , Yongchao Ma , Tong Xu , Guanglei Yang , Antonio Zandona , Tena Čadež , Zrinka Kovarik , Heidi Qunhui Xie , Li Xu , Bin Zhao","doi":"10.1016/j.cbi.2025.111722","DOIUrl":null,"url":null,"abstract":"<div><div>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 IC<sub>50</sub> 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 IC<sub>50</sub> values, eliminating the need for complex kinetic studies.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"421 ","pages":"Article 111722"},"PeriodicalIF":5.4000,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Simplified assessment of acetylcholinesterase inhibition by environmental organophosphorus pesticides\",\"authors\":\"Ruihong Zhu , Yiyun Liu , Yongchao Ma , Tong Xu , Guanglei Yang , Antonio Zandona , Tena Čadež , Zrinka Kovarik , Heidi Qunhui Xie , Li Xu , Bin Zhao\",\"doi\":\"10.1016/j.cbi.2025.111722\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>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 IC<sub>50</sub> 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 IC<sub>50</sub> values, eliminating the need for complex kinetic studies.</div></div>\",\"PeriodicalId\":274,\"journal\":{\"name\":\"Chemico-Biological Interactions\",\"volume\":\"421 \",\"pages\":\"Article 111722\"},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2025-09-04\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Chemico-Biological Interactions\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0009279725003527\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chemico-Biological Interactions","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0009279725003527","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Simplified assessment of acetylcholinesterase inhibition by environmental organophosphorus pesticides
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.
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.