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两种新型斑马鱼模型的比较分析:低 CYP1A 表达系和 CYP1A 敲除系在多环芳烃暴露下的表现。

Comparative analysis of two new zebrafish models: The cyp1a low-expression line and cyp1a knockout line under PAHs exposure.

机构信息

College of Marine Sciences, South China Agricultural University, Guangzhou, Guangdong, 510642, China; Guangdong Laboratory for Lingnan Modern Agriculture, South China Agricultural University, Guangzhou, 510642, China; Joint Laboratory of Guangdong Province and Hong Kong Region on Marine Bioresource Conservation and Exploitation, South China Agricultural University, Guangzhou, 510642, China.

College of Biological and Agricultural Engineering, Weifang University, Weifang 261061, Shandong, China.

出版信息

Gene. 2023 Jun 15;869:147391. doi: 10.1016/j.gene.2023.147391. Epub 2023 Mar 24.

Abstract

Cytochrome P450 1 (CYP1) is an important enzyme family involved in the metabolism of pollutants, and used as a biomarker to monitor environmental pollution. In this study, a fluorescence-labeled cyp1a zebrafish line, named as KI (cyp1a-T2A-mCherry) (KICM), was originally constructed to monitor dioxin-like compounds in the environment. However, the cyp1a gene expression in the KICM line was inhibited by the fluorescence labeling, thus leading to a significantly increased sensitivity of KICM zebrafish line to PAHs. Then, a cyp1a knockout zebrafish line, named KOC, were constructed for comparative analysis with the cyp1a low-expression line. Interestingly, knockout of the cyp1a gene did not increase the sensitivity of zebrafish to PAHs as significantly as the cyp1a low-expression line. So, the expression levels of related genes in the aryl hydrocarbon receptor pathway were analyzed and the results showed that the expression level of cyp1b in KOC group was significantly higher than that of wild type and KICM under the same PAH exposure. This indicated that the effect of losing cyp1a was compensated by inducing expression of cyp1b. In conclusion, two new zebrafish models including cyp1a low-expression line and cyp1a knockout line were constructed in this study, which may provide a convenient model for subsequent studies on the toxicity mechanism of PAHs and the role of cyp1a in detoxification.

摘要

细胞色素 P4501(CYP1)是参与污染物代谢的重要酶家族,被用作监测环境污染的生物标志物。在这项研究中,构建了一种荧光标记的 cyp1a 斑马鱼系,命名为 KI(cyp1a-T2A-mCherry)(KICM),最初用于监测环境中的二恶英类化合物。然而,荧光标记抑制了 KICM 系中 cyp1a 基因的表达,从而导致 KICM 斑马鱼系对多环芳烃的敏感性显著增加。然后,构建了 cyp1a 敲除斑马鱼系,命名为 KOC,用于与 cyp1a 低表达系进行比较分析。有趣的是,cyp1a 基因敲除并没有像 cyp1a 低表达系那样显著增加斑马鱼对多环芳烃的敏感性。因此,分析了芳基烃受体途径中相关基因的表达水平,结果表明,在相同的多环芳烃暴露下,KOC 组中 cyp1b 的表达水平明显高于野生型和 KICM。这表明 cyp1a 的缺失效应通过诱导 cyp1b 的表达得到了补偿。总之,本研究构建了两种新的斑马鱼模型,包括 cyp1a 低表达系和 cyp1a 敲除系,这可能为后续研究多环芳烃的毒性机制以及 cyp1a 在解毒中的作用提供便利的模型。

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