淡色库蚊抗吡丙醚及敏感种群解毒酶活性及其mRNA的表达差异  

Activity and gene expression of detoxifying enzymes from pyriproxyfen resistant population and susceptible population of Culex pipiens pallens

作　　者：刘洪霞[1] 刘曜[1] 冷培恩[1] LIU Hong-xia;LIU Yao;LENG Pei-en(Shanghai Municipal Center for Disease Control and Prevention , Shanghai 200336, China)

机构地区：[1]上海市疾病预防控制中心,上海200336

出　　处：《中华卫生杀虫药械》2019年第2期173-176,共4页Chinese Journal of Hygienic Insecticides and Equipments

基　　金：上海市自然科学基金项目(编号:16ZR140666)

摘　　要：目的探讨2种解毒酶在淡色库蚊吡丙醚抗性中的作用及其相关基因mRNA表达量的差异。方法采用分光光度法测定羧酸酯酶(CarE)和谷胱甘肽硫转移酶(GSTs)的酶活性,采用实时荧光定量PCR(Real-time PCR)测定CarE及GSTs相关基因的相对表达量。结果淡色库蚊抗吡丙醚种群和敏感种群CarE和GSTs的最适底物分别为1-氯-2,4-二硝基苯(CDNB)和乙酸-2-萘酯(β-NA)。2个蚊虫种群CarE活力分别为3.998×10^(-2)△OD/(mg protein·min)和3.010×10^(-2)△OD/(mg protein·min),两者差异有统计学意义(t=5.486,P<0.05);GSTs酶活力分别为3.626×10^(-4)和2.737×10^(-4)nmol/(mg protein·min),两者差异有统计学意义(t=3.899,P<0.05)。2个种群EST-2、GST1、GST2基因mRNA相对表达量差异无统计学意义(P>0.05),EST-3基因mRNA相对表达量差异有统计学意义(P<0.05)。结论吡丙醚可能诱导淡色库蚊EST-3基因上调表达而增强了羧酸酯酶的解毒酶活性,GSTs酶活性升高与GST1和GST2基因没有关系。Objective To explore the function of two detoxifying enzyme(carboxylesterase and glutathione S-transferase) in pyriproxyfen-resistant population of Culex pipiens pallens and know the expression of related gene.Methods The activities of glutathione S-transferases(GSTs) and carboxylesterase were determined based on the methods of Habig(1976)and van Aspern(1962).The mRNA expression of related gene was measured by real-time RCR.Results The optimum substrates of carboxylesterase(CarE) and glutathione S-transferase(GSTs) from different populations were 1-Chloro-2,4-dinitrobenzene(CDNB)and 2-Naphthyl acetate(β-NA).The activities of CarE were 3.998×10^-2 and 3.010×10^-2△OD/(mg protein·min) respectively in the resistant and the susceptible population with significant difference(t=5.486,P<0.05).The activities of GSTs were 3.626×10-4 and 2.737×10^-4 nmol/(mg protein ·min) with significant difference(t=3.899,P<0.05).The relative expression levels of EST-2 gene,GST1 gene and GST2 gene in the two populations were little different with that of the susceptible population(P>0.05),while some difference between the two populations in relative expression of EST-3(P<0.05).Conclusion Pyriproxyfen may up-regulate the expression of EST-3 gene,and enhance the activity of CarE in Cx.pipiens pallens,and the expressions of GST1 and GST2 have little relationship with the increasing of GSTs activity.

关 键 词：淡色库蚊 吡丙醚 羧酸酯酶 谷胱甘肽硫转移酶 

分 类 号：R384.1[医药卫生—医学寄生虫学]