溴敌隆对斑马鱼早期发育阶段的神经毒性作用

吴元钊; 张安慧; 徐帆; 郑中堉; 应剑波; 王斌杰; 王继业; 姚伟宣

doi:10.11836/JEOM21576

溴敌隆对斑马鱼早期发育阶段的神经毒性作用

Neurodevelopmental toxicity of bromadiolone to early-staged zebrafish

摘要

摘要:
背景溴敌隆是国内外广泛使用的第二代抗凝血类杀鼠剂，在生命早期暴露会导致神经发育毒性，但对于神经发育的毒性机制作用尚不明确。
目的探究溴敌隆暴露导致斑马鱼胚胎神经发育毒性及可能的作用机制。
方法将斑马鱼胚胎随机分为4组，分别为对照组（二甲基亚砜）以及0.39、0.78、1.18 mg·L⁻¹溴敌隆暴露组。暴露时间为受精后4~120 h。暴露至24 h记录每分钟内斑马鱼胚胎自主抽动次数，至120 h记录斑马鱼幼鱼120 min内的运动情况，检测幼鱼体内的乙酰胆碱酯酶（AChE）活性，通过荧光定量PCR方法检测神经发育相关基因（elavl3、gap43、mbp、syn2a）表达情况。
结果与对照组相比，24 h时1.18 mg·L⁻¹溴敌隆暴露组的斑马鱼胚胎自主抽动次数下降40%（P<0.05）。120 h时，当溴敌隆暴露浓度达到0.78、1.18 mg·L⁻¹时，运动总距离相较于对照组分别下降60%、69%（P<0.05、P<0.01），运动总时间相较于对照组分别下降34%、65%（P<0.05、P<0.01）。与对照组相比，1.18 mg·L⁻¹暴露组AChE活性升高36%（P<0.05）。荧光定量PCR测试结果表明：与对照组相比，神经发育关键基因elavl3、syn2a和mbp的基因表达在1.18 mg·L⁻¹暴露组分别下调66%、69%、65%（P<0.01），gap43基因表达在溴敌隆浓度为0.78 mg·L⁻¹时上调56%（P<0.01），在浓度为1.18 mg·L⁻¹时，表达量下调34%（P<0.05）。
结论溴敌隆暴露抑制斑马鱼胚胎自主抽动、运动行为能力，斑马鱼神经发育相关基因表达下调，阻碍神经传导递质释放，从而对斑马鱼产生神经发育毒性作用。

Abstract:
Background Bromadiolone is the second-generation anticoagulant rodenticide widely used all over the world. Exposure to bromadiolone in early life stage can lead to neurodevelopmental toxicity, but its toxic mechanism of neurodevelopment is not clear so far.
Objective To investigate the developmental neurotoxicity and mechanism of bromadiolone to zebrafish embryos.
Methods Zebrafish embryos were randomly divided into four groups: a solvent control group (dimethylsulphoxide) and three bromadiolone exposure groups (0.39, 0.78, and 1.18 mg·L⁻¹). The exposure period was from 4 h to 120 h post-fertilization. The number of spontaneous movement per minute was recorded at 24 h post-treatment. The locomotor ability of zebrafish larvae and the activity of acetylcholinesterase (AChE) were tested at 120 h post-treatment. The relative expression levels of neurodevelopment-related genes (elavl3, gap43, mbp, and syn2a) were measured by fluorescence quantitative PCR.
Results Compared with the control group, the number of spontaneous movement per minute at 24 h decreased significantly in the 1.18 mg·L⁻¹ bromadiolone exposure group (P<0.05). Compared with the control group, the total distance travelled of the zebrafish larvae in the 0.78 and 1.18 mg·L⁻¹ bromadiolone exposure groups decreased by 60% and 69% respectively (P<0.05, P<0.01), and the total movement time decreased by 34% and 65% respectively (P<0.05, P<0.01). The AChE activity in the 1.18 mg·L⁻¹ bromadiolone exposure group increased by 36% when compared with the control group (P<0.05). The fluorescence quantitative PCR results showed that compared with the control group, the expression levels of neurodevelopment-related genes elavl3, syn2a, and mbp were significantly down-regulated by 66%, 69%, and 65% in the 1.18 mg·L⁻¹ bromadiolone exposure group respectively (P<0.01), the expression level of gap43 was up-regulated by 56% in the 0.78 mg·L⁻¹ bromadiolone exposure group (P<0.01) and down-regulated by 34% in the 1.18 mg·L⁻¹ bromadiolone exposure group (P<0.05).
Conclusion Bromadiolone exposure could inhibit spontaneous movement and locomotive behavior, down-regulate the expression levels of neurodevelopment-related genes, hinder the release of neurotransmitters, and result in neurodevelopmental toxicity in the early-staged zebrafish.

HTML全文

参考文献(26)

施引文献

补充资料

审稿意见

勘误/撤稿