AHR2 morpholino knockdown reduces the toxicity of total particulate matter to zebrafish embryos
AHR2 morpholino knockdown reduces the toxicity of total particulate matter to zebrafish embryos
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AHR2吗啉敲除降低了总颗粒物对斑马鱼胚胎的毒性
DOI：
10.1016/j.taap.2016.08.024
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发表时间：
2016-10-15
影响因子：
3.8
通讯作者：
Di Giulio, Richard T.
中科院分区：
文献类型：
--
作者：
Massarsky, Andrey;Bone, Audrey J.;Di Giulio, Richard T.
斑马鱼胚胎被提议作为研究香烟烟雾对早期发育影响的“桥梁模型”。研究表明，总颗粒物（TPM）暴露对斑马鱼的生长发育有不良影响，抗氧化剂和芳香烃受体（AHR）途径在其中起重要作用。本研究探讨了这两种途径在介导TPM毒性中的作用。该研究包括四个实验。在实验I中，斑马鱼胚胎从受精后6小时（hpf），直到96 hpf暴露于TPM0.5和TPM1.0（对应于0.5和1.0 μ g/mL当量尼古丁单位）的存在或不存在的抗氧化剂（N-乙酰半胱氨酸/NAC）或促氧化剂（丁硫酰亚胺/BSO）。在实验II中，TPM曝光进行了显微注射与核因子红细胞2相关因子2（Nrf 2），AHR 2，细胞色素P450 1A（CYP 1A），或CYP 1B 1吗啉的胚胎，畸形进行了评估。在实验III中，胚胎暴露于TPM，并在24，48，72和96 hpf收集胚胎/幼虫，以评估与抗氧化剂和AHR途径相关的几个基因。最后，实验IV评估了暴露于TPM后CYP 1A和CYP 1B 1的活性和蛋白水平。我们证明TPM诱导的畸形的发生率通常不受NAC/BSO治疗或Nrf 2敲低的影响。相反，AHR 2敲低降低，而CYP 1A或CYP 1B 1敲低升高某些畸形的发生率。此外，如基因表达所示，TPM暴露诱导了AHR途径而非抗氧化途径，进一步证明了其在介导TPM毒性中的重要性。(C)版权所有© 2016 Elsevier Inc.
The zebrafish embryo has been proposed as a 'bridge model' to study the effects of cigarette smoke on early development. Previous studies showed that exposure to total particulate matter (TPM) led to adverse effects in developing zebrafish, and suggested that the antioxidant and aryl hydrocarbon receptor (AHR) pathways play important roles. This study investigated the roles of these two pathways in mediating TPM toxicity. The study consisted of four experiments. In experiment I, zebrafish embryos were exposed from 6 h post fertilization (hpf) until 96 hpf to TPM0.5 and TPM1.0 (corresponding to 0.5 and 1.0 mu g/mL equi-nicotine units) in the presence or absence of an antioxidant (N-acetyl cysteine/NAC) or a pro-oxidant (buthionine sulfoximine/BSO). In experiment II, TPM exposures were performed in embryos that were microinjected with nuclear factor erythroid 2-related factor 2 (Nrf2), AHR2, cytochrome P450 1A (CYP1A), or CYP1B1 morpholinos, and deformities were assessed. In experiment III, embryos were exposed to TPM, and embryos/larvae were collected at 24, 48, 72, and 96 hpf to assess several genes associated with the antioxidant and AHR pathways. Lastly, experiment IV assessed the activity and protein levels of CYP1A and CYP1B1 after exposure to TPM. We demonstrate that the incidence of TPM-induced deformities was generally not affected by NAC/BSO treatments or Nrf2 knockdown. In contrast, AHR2 knockdown reduced, while CYP1A or CYP1B1 knockdowns elevated the incidence of some deformities. Moreover, as shown by gene expression the AHR pathway, but not the antioxidant pathway, was induced in response to TPM exposure, providing further evidence for its importance in mediating TPM toxicity. (C) 2016 Elsevier Inc All rights reserved.