Laboratório de Enzimologia-LABENZ, Departamento de Bioquímica and Laboratório de Imunopatologia Keizo Asami, LIKA Universidade Federal de Pernambuco, Recife, PE, Brazil.
Hum Exp Toxicol. 2013 Aug;32(8):837-45. doi: 10.1177/0960327112468906. Epub 2013 Apr 30.
A method of extracting membranes from red blood cells (RBCs) is described, which were in turn used to assay acetylcholinesterase (AChE) activity. The evidence for the enzyme activity was established by selective inhibition using 1,5-bis(4-allyldimethylammoniumphenyl) pentan-3-one dibromide, tetraisopropyl pyrophosphoramide and neostigmine. Blood samples were exposed to three organophosphorus (dichlorvos, chlorpyrifos and diazinon) and two carbamate (carbaryl and carbofuran) pesticides. Afterwards AChE activities in RBC membranes were determined. The concentrations capable to inhibit the enzyme activity by 50% (IC₅₀) for the pesticides were 10.66 µM (dichlorvos), 21.42 µM (chlorpyrifos), 109.98 µM (carbaryl) and 5.44 µM (carbofuran). The results related to 20% enzyme inhibition (level used in the estimation of threshold limits for anticholinesterase compounds) were below those acceptable daily intake values enacted by relevant national and international regulations. These results suggest that the proposed AChE extraction from RBC and assay could be a suitable method for monitoring occupational exposure to pesticides.
一种从红细胞（RBC）中提取膜的方法被描述，然后用该膜来测定乙酰胆碱酯酶（AChE）活性。通过使用 1,5-双（4-丙烯基二甲基铵苯基）戊烷-3-酮二溴化物、四异丙基焦磷酸酰胺和新斯的明进行选择性抑制，建立了酶活性的证据。血液样本暴露于三种有机磷（二氯松、毒死蜱和二嗪农）和两种氨基甲酸酯（西维因和呋喃丹）农药中。之后，测定 RBC 膜中 AChE 的活性。能够抑制酶活性 50%（IC₅₀）的农药浓度分别为 10.66µM（二氯松）、21.42µM（毒死蜱）、109.98µM（西维因）和 5.44µM（呋喃丹）。与 20%酶抑制（用于估计抗胆碱酯酶化合物的阈值限制）相关的结果低于相关国家和国际法规规定的可接受日摄入量值。这些结果表明，从 RBC 中提取 AChE 并进行测定的方法可能是监测职业暴露于农药的一种合适方法。
