双酚AP和双酚AF对斑马鱼的早期神经发育毒性作用研究

Neurodevelopmental toxicities of bisphenol AP and bisphenol AF in early life of zebrafish

  • 摘要:
    目的 双酚AP(bisphenol AP,BPAP)和双酚AF(bisphenol AF,BPAF)大规模的生产和使用,对水生生态环境具有潜在威胁。关于BPAP和BPAF的神经毒性研究甚少,本文以斑马鱼作为模式动物,研究BPAP和BPAF对斑马鱼的早期神经发育毒性作用。

    方法 本研究选取斑马鱼幼鱼作为实验对象,将斑马鱼胚胎随机分为5组,以BPAP(20μg/L和200 μg/L)和BPAF(20 μg/L和200 μg/L)进行染毒,染毒时间为受精后4 h至受精后144 h。通过斑马鱼的运动行为学测试,原位杂交和荧光定量PCR方法来阐述BPAP和BPAF的早期神经发育毒性作用及其可能的作用机制。

    结果 行为学测试结果显示,与对照组相比,染毒组的斑马鱼幼鱼的运动总距离和速度均明显下降,热图显示染毒组幼鱼有过多呆滞不动的现象,表明BPAP、BPAF均抑制了斑马鱼幼鱼的移动距离和速度。荧光定量PCR结果可见,与对照组相比,BPAP(20 μg/L和200 μg/L)和BPAF(20 μg/L和200 μg/L)都抑制了斑马鱼神经发育关键基因(mbpsyn2a)的表达(P < 0.001),同时原位杂交结果也验证了基因的结果。

    结论 初步认为BPAF和BPAP对斑马鱼具有潜在的神经发育毒性作用。

     

    Abstract:
    Objective Mass production and utilization of bisphenol AP (BPAP) and bisphenol AF (BPAF) has posed potential ecological risk to aquatic ecosystem. However, few studies have focused on the neurotoxicities of BPAP and BPAF. This study investigates the developmental neurotoxicities of BPAP and BPAF in the early life of zebrafish.

    Methods We selected zebrafish larvae as experimental subjects. Zebrafish embryos were randomly divided into five groups. BPAP (20 μg/L and 200 μg/L) and BPAF (20 μg/L and 200 μg/L) were administered with the designed drug exposure profile from 4 h to 144 h after fertilization. The early developmental neurotoxicities of BPAP and BPAF and their possible mechanisms were discussed by locomotion behavior test, in situ hybridization, and fluorescence quantitative PCR.

    Results Compared with the control group, the total moving distance and the speed of the zebrafish larvae in the exposed groups were significantly decreased, and the thermo gram showed that the larvae in the exposed groups were sluggish, indicating that both BPAP and BPAF inhibited the moving distance and speed of zebrafish larvae. The results of fluorescence quantitative PCR showed that compared with the control group, BPAP (20 μg/L and 200 μg/L) and BPAF (20 μg/L and 200μg/L) significantly inhibited the expressions of key genes (mbp and syn2a) involved in zebrafish neurodevelopment. The results of in situ hybridization also verified the results of the genes.

    Conclusion The findings tentatively suggest that BPAF and BPAP may have potential developmental neurotoxic effects on zebrafish.

     

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