阻燃剂磷酸三丁酯对斑马鱼早期发育的影响  

Effects of flame retardant tributyl phosphate on early development of zebrafish

作　　者：李瑶 朱晶颖 李尧 陈丽梅 朱鹏飞[1] 丁新良[1] 周伟杰[1] LI Yao;ZHU Jingying;CHEN Limei;ZHU Pengfei;DING Xinliang;ZHOU Weijie(Wuxi Center for Disease Control and Prevention(Wuxi Medical Center,Nanjing Medical University),Wuxi,Jiangsu 214400,China)

机构地区：[1]无锡市疾病预防控制中心(南京医科大学无锡医学中心),江苏无锡214400

出　　处：《环境与职业医学》2024年第12期1376-1383,共8页Journal of Environmental and Occupational Medicine

基　　金：南京医科大学无锡医学中心2023年“揭榜挂帅”项目(WMCJ202302);无锡市水质健康研究队列建设项目(WMCC202318)。

摘　　要：[背景]磷酸三丁酯(TBP)作为一种有机磷酸酯阻燃剂被广泛应用,然而TBP在低浓度暴露下对水生生物毒性研究有限。[目的]以斑马鱼作为模式动物,探讨阻燃剂TBP对斑马鱼早期发育的影响。[方法]将受精后2 h(2 hpf)的斑马鱼胚胎随机分为4组,分别为0.01%二甲基亚砜(DMSO)对照组和TBP染毒组(0.02、0.2、2μg·L^(-1))。染毒时间为2 hpf至120 hpf,分别观察斑马鱼胚胎72 hpf孵化率、畸形率、心率和体长,24~29 hpf卷尾频率、96 hpf运动能力和120 hpf存活率。染毒结束后利用酶联免疫法检测幼鱼全身三碘甲状腺原氨酸(T3)及甲状腺素(T4)含量,采用实时荧光定量聚合酶链式反应(q-PCR)法检测下丘脑-垂体-甲状腺轴(HPT)和神经发育相关基因的表达水平。[结果]TBP染毒组斑马鱼胚胎均出现心率下降(P<0.001),0.02、2μg·L^(-1)染毒组的存活率下降(P<0.05),2μg·L^(-1)染毒组畸形率上升(P<0.05),主要表现为心包水肿。各组斑马鱼胚胎卷尾频率在25 hpf达到最高,各染毒组的卷尾频率低于对照组(P<0.001)。运动行为实验中,暗周期0.02、0.2μg·L^(-1)染毒组斑马鱼游泳速度下降(P<0.05),在光周期0.2、2μg·L^(-1)染毒组斑马鱼游泳速度明显下降(P<0.05)。与对照组相比,0.2μg·L^(-1)染毒组幼鱼T3水平上升(P<0.05)。q-PCR结果表明,染毒组HPT轴相关基因甲状腺激素受体(trα、trβ)、甲状腺球蛋白(tg)、钠碘共转运体(nis)表达水平下调,甲状腺转运蛋白(ttr)在0.02μg·L^(-1)染毒组上调,碘甲腺原氨酸脱碘酶2(dio2)在0.02μg·L^(-1)染毒组下调(P<0.05);染毒组神经发育相关基因乙酰胆碱酯酶(ache)下调,髓鞘碱性蛋白(mbp)和ELAV样神经元特异性RNA结合蛋白3(elavl3)在0.02μg·L^(-1)染毒组也下调(P<0.05)。[结论]TBP暴露可导致斑马鱼早期发育异常,表现为孵化期和早幼期的发育毒性、甲状腺内分泌破坏和神经毒性。[Background]Tributyl phosphate(TBP)is widely used as an organophosphate flame retardant.However,there are limited studies on the toxicity of TBP to aquatic organisms at low levels of exposure.[Objective]To investigate the effects of TBP on early development of zebrafish(Danio rerio).[Methods]Zebrafish embryos were randomly divided into four groups at 2 h post-fertilisation(2 hpf),namely,the 0.01%dimethyl sulfoxide(DMSO)control group and TBP exposure groups(0.02,0.2 and 2μg·L^(−1)).The exposure time was from 2 hpf to 120 hpf and the hatching rate,malformation rate,heart rate and body length of zebrafish embryos at 72 hpf,the frequency of tail curling at 24-29 hpf,the locomotor ability at 96 hpf and the survival rate at 120 hpf were evaluated,respectively.The whole-body triiodothyronine(T3)and tetraiodothyronine(T4)levels of juvenile fish were measured by enzyme immunoassay at the end of the infection,and the expression levels of hypothalamic-pituitary-thyroid axis(HPT)and neurodevelopmental-related genes were detected by quantitative real-time PCR(q-PCR).[Results]The heart rates of zebrafish embryos were significantly decreased in all TBP-treated groups(P<0.001),the survival rates of the 0.02 and 2μg·L^(−1) TBP groups were significantly decreased(P<0.05),and the malformation rate of the 2μg·L^(−1) treated group was significantly increased(P<0.05),which was mainly manifested by pericardial oedema.The frequency of tail curling of zebrafish embryos in all groups reached the highest at 25 hpf,which was significantly lower(P<0.001)in all exposure groups than in the control group(P<0.001).In the locomotor behaviour experiments,the swimming speed of zebrafish larvae in the dark cycle was significantly decreased in the 0.02 and 0.2μg·L^(−1) TBP groups(P<0.05),and similar results were found for the light cycle in the 0.2 and 2μg·L^(−1) TBP groups(P<0.05).Compared with the control group,the T3 level of zebrafish juveniles in the 0.2μg·L^(−1) TBP group increased significantly(P<0.05).The q-PCR results

关 键 词：有机磷酸酯 磷酸三丁酯 斑马鱼 内分泌干扰 神经毒性 发育毒性 

分 类 号：R114[医药卫生—卫生毒理学]