芳香烃受体介导双酚A致斑马鱼胚胎发育毒性  

Aryl hydrocarbon receptor-mediated developmental toxicity in zebrafish embryos induced by bisphenol A

作　　者：王伟 何梦婷 许洁 张洁[1] WANG Wei;HE Mengting;XU Jie;ZHANG Jie(Department of Health Toxicology,School of Public Health,Medical College of Soochow University,Suzhou,Jiangsu 215123,China)

机构地区：[1]苏州大学医学部公共卫生学院卫生毒理学教研室,江苏苏州215123

出　　处：《环境与职业医学》2021年第4期342-349,共8页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金项目(81673203)。

摘　　要：[背景]双酚A(BPA)在生命早期暴露会导致发育毒性,芳香烃受体(AhR)能够调控各种发育和生理功能,研究发现BPA可使其激活。[目的]以斑马鱼作为模式动物,研究AhR在BPA所致斑马鱼胚胎发育毒性中的作用。[方法]将斑马鱼胚胎随机分为8组进行染毒,分别为DMSO对照组、AhR抑制剂CH223191(CH)对照组、BPA暴露组(1、5、25μmol·L^(-1))、CH+BPA(1、5、25μmol·L^(-1))组。染毒时间为受精后2 h(2 hpf)至受精后96 h(96 hpf),分别观察斑马鱼胚胎的死亡率、孵化率、畸形率、心率、体长及运动能力。通过荧光定量PCR方法检测AhR相关基因(ahr2、cyp1a1、cyp1b1)、心脏发育关键基因(nkx2.5、sox9b)、神经发育关键基因(elavl3、gfap、mbp、syn2a)、氧化应激相关基因(nrf2、sod1/2、ho1、nqo1)的表达。[结果]与DMSO对照组相比,25μmol·L^(-1) BPA暴露组斑马鱼胚胎出现孵化延迟、心率下降、畸形率升高,5μmol·L^(-1)和25μmol·L^(-1) BPA暴露组出现死亡率增加、体长缩短以及运动距离下降的现象(P<0.05)。与相应BPA暴露组相比,CH+25μmol·L^(-1) BPA组斑马鱼的心率、畸形率、运动能力和CH+5μmol·L^(-1) BPA组及CH+25μmol·L^(-1) BPA组的体长均有所恢复(P<0.05)。但孵化率和死亡率并未受CH影响。荧光定量PCR结果表明,BPA组AhR相关基因(ahr2、cyp1a1和cyp1b1)的表达上调(P<0.05);CH联合处理后能降低各BPA组ahr2、cyp1a1基因的过表达及25μmol·L^(-1) BPA组cyp1b1的过表达(P<0.05)。心脏发育相关基因(nkx2.5和sox9b)的表达分别在25μmol·L^(-1)和5、25μmol·L^(-1) BPA组出现下调(P<0.05);CH联合处理后上述基因的低表达恢复(P<0.05)。神经发育相关基因elavl3在25μmol·L^(-1) BPA组出现下调,gfap和syn2a在5、25μmol·L^(-1) BPA组出现下调,mbp在各BPA组均出现下调(P<0.05);CH联合处理后上述基因的低表达恢复(P<0.05)。BPA暴露后氧化应激相关基因的表达也出现改变,其中nrf2在25μmol·L^(-1) BPA组出�[Background]Exposure to bisphenol A(BPA)in the early life period can induce developmental toxicity.Aryl hydrocarbon receptor(AhR)can regulate various developmental and physiological functions,and can be activated by BPA.[Objective]This experiment uses zebrafish as a model animal to study the role of AhR in the developmental toxicity of zebrafish embryos induced by BPA.[Methods]Zebrafish embryos were randomly divided into eight groups:DMSO control group,AhR inhibitor CH223191(CH)control group,BPA exposure groups(1,5,and 25μmol·L^(-1)),and CH+BPA(1,5,and 25μmol·L^(-1))groups.The exposure period was from 2 h post-fertilization(2 hpf)to 96 h post-fertilization(96 hpf).The mortality,hatching rate,malformation rate,heart rate,body length,and locomotor ability of zebrafish embryos were evaluated.The expressions of AhR-related genes(ahr2,cyp1 a1,and cyp1 b1),cardiac development-related genes(nkx2.5 and sox9 b),and neurodevelopment-related genes(elavl3,gfap,mbp,and syn2 a),and reactive oxygen species-related genes(nrf2,sod1,sod2,ho1,and nqo1)were detected by fluorescence quantitative PCR.[Results]Compared with the DMSO control group,the zebrafish embryos in the 25μmol·L^(-1) BPA group had delayed hatching,decreased heart rate,and elevated malformation rate,and the 5μmol·L^(-1) and 25μmol·L^(-1) groups showed shortened body length,increased mortality,and decreased movement distance(P<0.05).Compared with corresponding BPA exposure groups,the heart rate,malformation rate,and locomotor ability of the zebrafish in the CH+25μmol·L^(-1) BPA group,and the body length of the CH+5μmol·L^(-1) BPA group and the CH+25μmol·L^(-1) BPA group all recovered(P<0.05).However,the hatching rate and mortality rate were not affected by CH.The results of fluorescence quantitative PCR showed that Ah R-related genes(ahr2,cyp1 a1,and cyp1 b1)were up-regulated in the BPA groups(P<0.05),and CH combined treatment reduced the overexpression of ahr2 and cyp1 a1 in each BPA group and cyp1 b1 in the 25μmol·L^(-1) BPA group(P<0.05).The exp

关 键 词：双酚A 芳香烃受体 氧化应激 斑马鱼 发育毒性 

分 类 号：R114[医药卫生—卫生毒理学]