TBBPA-DHEE暴露对斑马鱼的神经毒性及机制研究  

Neurotoxicity and mechanisms of TBBPA-DHEE exposure in zebrafish

作　　者：徐彤 冯伟伟 丁阳阳 钱显 陈瑶 茆广华 仰榴青[2] 吴向阳 XU Tong;FENG Weiwei;DING Yangyang;QIAN Xian;CHEN Yao;MAO Guanghua;YANG Liuqing;WU Xiangyang(School of Environment and Safety Engineering;School of Chemistry and Chemical Engineering,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学环境与安全工程学院,江苏镇江212013 [2]江苏大学化工学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2021年第3期227-233,263,共8页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金面上项目(21976072)。

摘　　要：目的:研究四溴双酚A双(2-羟基乙基)醚[TBBPA bis(2-hydroxyetyl)ether,TBBPA-DHEE]对斑马鱼胚胎的神经系统影响及其机制。方法:将斑马鱼胚胎随机分为5组,分别为空白对照组、二甲基亚砜组、TBBPA-DHEE不同剂量组(0.086、0.860、8.600 mg/L),暴露5 d,观察其死亡率、孵化率、畸形率、心跳频率及体长;通过行为学实验分析胚胎自主运动情况及明暗刺激下仔鱼的游泳距离及累计持续移动时间;采用试剂盒检测仔鱼体内超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)活力及丙二醛含量,检测Ca^(2+)与γ-氨基丁酸含量;采用实时荧光定量PCR法测定Ca^(2+)信号通路相关mRNA表达。结果:(1)TBBPA-DHEE暴露显著增加斑马鱼胚胎/仔鱼的死亡率和畸形率;抑制其孵化;导致斑马鱼心跳频率和体长显著下降。(2)TBBPA-DHEE暴露显著降低斑马鱼胚胎自主运动次数;0.860和8.600 mg/L TBBPA-DHEE暴露显著降低黑暗条件下仔鱼游泳距离及累计持续移动时间。(3)TBBPA-DHEE暴露显著升高斑马鱼仔鱼体内SOD、GPx活力及丙二醛含量,诱导Ca^(2+)含量上升且显著抑制γ-氨基丁酸含量。(4)TBBPA-DHEE暴露显著上调Ca^(2+)信号通路中Cacna1ab、Atp2a1、Atp2a1l、Camk1ga、Ppp3ca、Plcd3a和Prkca mRNA表达。结论:TBBPA-DHEE可对斑马鱼胚胎产生神经毒性,可能通过增强内源抗氧化酶活性、诱导Ca^(2+)信号通路相关mRNA表达等起作用。Objective:To study the neurotoxicity and mechanisms of tetrabromobisphenol A bis(2-hydroxyetyl)ether(TBBPA-DHEE)on zebrafish.Methods:Zebrafish embryos were randomly divi-ded into five groups:control group,dimethyl sulfoxide(DMSO)group,TBBPA-DHEE exposure groups(0.086,0.860 and 8.600 mg/L).After exposure for five days,the mortality,hatching rate,malformation rate,heart rate and body length were observed.Behavioral experiments were used to analyze the spontaneous movement of embryos and the swimming distance and cumulative duration of larvae under light and dark stimulation.The activities of superoxide dismutase(SOD)and glutathione peroxidase(GPx),the contents of malondialdehyde,Ca^(2+)andγ-aminobutyric acid in larvae were detected by the assay kits.The mRNA expressions of calcium signaling pathway were detected by quantitative real-time PCR method.Results:(1)TBBPA DHEE exposure could greatly increase the mortality and malformation of zebrafish embryos/larval,and inhibit the incubation of embryos.The heart rate and body length were decreased remarkably.(2)TBBPA-DHEE exposure could significantly reduce the number of spontaneous movement of zebrafish embryos;0.860 and 8.600 mg/L TBBPA-DHEE exposure significantly reduced the swimming distances and cumulative mobile durations under dark condition.(3)TBBPA-DHEE exposure could increase the activities of SOD and GPx and the content of malondialdehyde.The content of Ca^(2+)has been increased and the content of gamma-aminobutyric acid has been decreased significantly after TBBPA-DHEE exposure.(4)TBBPA-DHEE exposure could significantly up-regulate the mRNA expressions of Cacna1ab,Atp2a1,Atp2a1l,Camk1ga,Ppp3ca,Plcd3a and Prkca in calcium signaling pathways.Conclusion:TBBPA-DHEE could induce the neurotoxicity of zebrafish embryos by enhancing the activities of endogenous antioxidant enzymes and inducing the expressions of related mRNA in calcium signaling pathway.

关 键 词：四溴双酚A衍生物 斑马鱼胚胎 氧化应激 Ca^(2+)信号通路 神经毒性 

分 类 号：R994[医药卫生—毒理学]