拟除虫菊酯对人羧酸酯酶2的抑制作用及动力学研究  

Inhibitory effect and kinetics analysis of pyrethroids on human carboxylesterase 2

作　　者：杜佐 孙珂钰 梁佳鸿 龚家敏 莫奉君 张星璐 矣肖镭 DU Zuo;SUN Ke-yu;LIANG Jia-hong;GONG Jia-min;MO Feng-jun;ZHANG Xing-lu;YI Xiao-lei(School of Public Health,North Sichuan Medical College,Nanchong,Sichuan 637100,China;School of Clinical Medicine,North Sichuan Medical College,Nanchong,Sichuan 637100,China;Chongqing Qjiang District for Disease Control and Prevention,Chongqing 401420,China)

机构地区：[1]川北医学院公共卫生学院,四川南充637100 [2]川北医学院临床医学院,四川南充637100 [3]重庆市綦江区疾病预防控制中心,重庆401420

出　　处：《毒理学杂志》2024年第5期344-351,共8页Journal of Toxicology

基　　金：川北医学院博士科研启动基金(CBY20-QD02);南充市市校科技战略合作专项资金项目(20SXQT0318)。

摘　　要：目的 研究拟除虫菊酯对人羧酸酯酶2(human carboxylesterase 2, hCE2)的抑制作用并测定相关抑制动力学参数。方法 以荧光素二乙酸酯(fluorescein diacetate, FD)作为hCE2的特异性底物,构建人肝微粒体(HLMs)催化反应的体外孵育体系,运用超高效液相色谱(UPLC)检测12种拟除虫菊酯(联苯菊酯、氯烯炔菊酯、甲醚菊酯、氯菊酯、氯氟氰菊酯、氯氰菊酯、溴氰菊酯、甲氰菊酯、氰戊菊酯、氟氰戊菊酯、三氟氯氰菊酯和氟胺氰菊酯)对hCE2的抑制作用。通过抑制动力学实验测定拟除虫菊酯对hCE2的半数抑制浓度(IC50)、抑制动力学类型和抑制动力学常数(Ki),并通过分子对接研究其分子间相互作用。结果 12种拟除虫菊酯(100μmol/L)均对hCE2产生明显的抑制作用,其中氰戊菊酯、氟氰戊菊酯和氟胺氰菊酯能够对hCE2产生强烈的抑制作用,抑制率大于80%(t值分别为15.87,17.65和12.96,P<0.05)。氰戊菊酯、氟氰戊菊酯和氟胺氰菊酯对hCE2的IC50分别为0.62,42.74和42.30μmol/L,抑制类型均为非竞争性抑制,Ki分别为24.30、6.36和24.70μmol/L。体外-体内外推(IVIVE)结果表明氟氰戊菊酯可能在体内抑制hCE2的催化代谢。分子对接结果表明拟除虫菊酯能够通过氢键和疏水作用与hCE2分子结合。结论 拟除虫菊酯能够对hCE2产生抑制作用,可能通过影响hCE2的催化反应产生相应毒性作用。Objective To investigate the inhibitory effect and kinetics parameters of pyrethroids on human carboxylesterase 2(hCE2).Methods Human liver microsomes(HLMs)catalysed hydrolysis of fluorescein diacetate(FD)was used as the probe reaction to detect the activity of hCE2.The residual activity of hCE2 was detected by ultra-high performance liquid chromatography(UPLC)after the intervention with 12 kinds of pyrethroids(bifenthrin,chlorempenthrin,methothrin,permethrin,cyhalothrin,cypermethrin,deltamethrin,fenpropathrin,fenvalerate,flucythrinate,lambda-cyhalothrin and tau-fluvalinate).Inhibition kinetics analysis was carried out to detect the half inhibitory concentration(IC50),inhibition type and kinetics parameter(Ki)of pyrethroids on hCE2.Molecular docking was performed to analyse the ligand-enzyme interaction.Results The result showed that all of 12 pyrethroids(100μmol/L)exhibited significant inhibition towards hCE2,among which fenvalerate,flucythrinate and tau-fluvalinate exhibited strong inhibition with inhibitory rates more than 80%(t=15.87,17.65 and 12.96,respectively,P<0.05).Based on the kinetics analysis,the IC50 values of fenvalerate,flucythrinate and tau-fluvalinate on hCE2 were calculated to be 0.62,42.74 and 42.30μmol/L,respectively,and the inhibition types were all non-competitive,with the Ki values being 24.30,6.36 and 24.70μmol/L,respectively.In vitro-in vivo extrapolation(IVIVE)indicated that flucythrinate might inhibit the metabolic catalysis in vivo by inhibiting hCE2.Molecular docking result showed that hydrogen bonds and hydrophobic contacts contributed to the interaction of pyrethroids and hCE2.Conclusion nPyrethroids exhibited inhibitory effect on hCE2,which might induce corresponding toxic effects by influencing the catalytic reaction of hCE2.

关 键 词：拟除虫菊酯 人羧酸酯酶2 酶抑制作用 抑制动力学参数 分子对接 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]