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黑鮶鱼细胞色素 P450 1 酶:苯并[a]芘暴露后的分子特征和表达模式。

Cytochrome P450 1 enzymes in black rockfish, Sebastes schlegelii: Molecular characterization and expression patterns after exposure to benzo[a]pyrene.

机构信息

Department of Aquatic Life Medicine, Pukyong National University, Busan 48513, South Korea; Pathology Division, National Instititue of Fisheries Science, Busan 46083, South Korea.

Department of Aquatic Life Medicine, Pukyong National University, Busan 48513, South Korea.

出版信息

Aquat Toxicol. 2020 Sep;226:105566. doi: 10.1016/j.aquatox.2020.105566. Epub 2020 Jul 8.

Abstract

Cytochrome P450 (CYP) enzymes play important roles in the detoxification and bioactivation of environmental contaminants and are involved in the responses to pollution in fish. In this study, we cloned four new CYP1 genes, CYP1A, CYP1B, CYP1C1, and CYP1C2, from black rockfish (Sebastes schlegelii), a dominant and economically important fish species in Korea. This species is at a significant risk of exposure to petrohydrocarbons, such as benzo[a]pyrene(B[a]P), due to frequent oil spills along the Korean coast. Quantitative PCR analysis of CYP1 gene transcription in 12 organs of the fish revealed tissue-specific expression patterns. CYP1A was significantly expressed in the liver, heart, kidneys, and muscle, and CYP1B was significantly expressed in the gills, muscle, and heart. CYP1C1 and CYP1C2 showed similar tissue expression patterns, with the highest levels in the muscle. Furthermore, exposure to an aryl hydrocarbon receptor (AHR) agonist, B[a]P, at 2, 20, and 200 mg/kg body weight showed significant dysregulation of the CYP1A, CYP1B, CYP1C1, and CYP1C2 expression levels in the gills, liver, kidneys, and spleen. The mRNA expression levels of CYP1A and CYP1B were upregulated by 450- and 17-fold, respectively, in the spleen. Compared with their levels in the control, CYP1C1 increased by 45-fold, while CYP1C2 remained unchanged in the gills, indicating differential effects of the polycyclic aromatic hydrocarbon on CYP1 expression in different fish organs. The results suggested that expression profiles of inducible CYP1 enzymes in S. schlegelii might be used as indicators for assessing aquatic contamination by AHR agonists. Determination of the basal and induced expression levels, as well as substrate specificity, of the four CYP1 enzymes may contribute to a better understanding of their roles in the metabolization of toxicants or drugs.

摘要

细胞色素 P450(CYP)酶在环境污染物的解毒和生物活化中发挥着重要作用,并参与鱼类对污染的反应。在这项研究中,我们从韩国主要的经济鱼类黑鲷(Sebastes schlegelii)中克隆了四个新的 CYP1 基因,CYP1A、CYP1B、CYP1C1 和 CYP1C2。由于韩国沿海经常发生石油泄漏,这种鱼类面临着接触石油烃如苯并[a]芘(B[a]P)等污染物的巨大风险。对鱼 12 种器官中 CYP1 基因转录的定量 PCR 分析显示出组织特异性表达模式。CYP1A 在肝脏、心脏、肾脏和肌肉中表达显著,CYP1B 在鳃、肌肉和心脏中表达显著。CYP1C1 和 CYP1C2 表现出相似的组织表达模式,在肌肉中的表达水平最高。此外,暴露于芳基烃受体(AHR)激动剂 B[a]P(2、20 和 200mg/kg 体重)导致鳃、肝脏、肾脏和脾脏中 CYP1A、CYP1B、CYP1C1 和 CYP1C2 的表达水平显著失调。在脾脏中,CYP1A 和 CYP1B 的 mRNA 表达水平分别上调了 450 倍和 17 倍。与对照组相比,CYP1C1 在鳃中增加了 45 倍,而 CYP1C2 保持不变,表明多环芳烃对不同鱼类器官中 CYP1 表达的影响不同。结果表明,S. schlegelii 中诱导型 CYP1 酶的表达谱可作为评估 AHR 激动剂引起的水生污染的指标。四种 CYP1 酶的基础和诱导表达水平以及底物特异性的测定可能有助于更好地理解它们在毒物或药物代谢中的作用。

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