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两种新型 CYP3A 同工酶在海洋贻贝贻贝中的鉴定及对镉和苯并[a]芘的响应。

Two novel CYP3A isoforms in marine mussel Mytilus coruscus: Identification and response to cadmium and benzo[a]pyrene.

机构信息

NationalEngineering Research Center of Marine Facilities Aquaculture, Marine Science and Technology College, Zhejiang Ocean University, Zhoushan, 316004, China.

Shaanxi Key Laboratory of Earth Surface System and Environmental Carrying Capacity, College of Urban and Environmental Sciences, Northwest University, Xi'an, 710127, China.

出版信息

Aquat Toxicol. 2019 Sep;214:105239. doi: 10.1016/j.aquatox.2019.105239. Epub 2019 Jul 2.

Abstract

CYP3A enzymes play a crucial role in metabolic clearance of a variety of xenobiotics. However, their genetic information and function remain unclear in molluscs. In the present study, two novel CYP3A genes i.e. McCYP3A-1 and McCYP3A-2 were identified and characterized from the thick shell mussel Mytilus coruscus, and their tissue distribution as well as the response to cadmium (Cd) and benzo[a]pyrene (B[α]P) exposure were addressed using real time quantitative RT-PCR (qRT-PCR) and erythromycin N-demethylase (ERND) assay. McCYP3A-1 and McCYP3A-2 possess typically domains of CYP family such as helix-C, helix-I, helix-K, PERF and the heme binding domain as well as the characteristic domains of CYP3s including six SRS motifs. McCYP3A-1 and McCYP3A-2 transcripts were constitutively expressed in all examined tissues with high expression level in digestive glands, hepatopancreas and gonads. Upon B[α]P exposure, McCYP3A-1 and McCYP3A-2 mRNA expression in digestive glands showed a pattern of up-regulation followed by down-regulation, while under Cd exposure, showed a time-dependent induction profile. In addition, ERND activity, generally used as an indicator of CYP3, increased in a time-dependent manner after exposure to Cd and B[α]P. These results collectively indicated that McCYP3A-1 and McCYP3A-2 are CYP3A family member and may play a potential role in metabolic clearance of xenobiotics. Meanwhile, the current results may provide some baseline data to support McCYP3A-1 and McCYP3A-2 as candidate biomarkers for monitoring of PAHs and heavy metal pollution.

摘要

CYP3A 酶在多种外源物质的代谢清除中起着至关重要的作用。然而,它们的遗传信息和功能在软体动物中仍然不清楚。本研究从厚壳贻贝 Mytilus coruscus 中鉴定并表征了两个新的 CYP3A 基因,即 McCYP3A-1 和 McCYP3A-2,并通过实时定量 RT-PCR(qRT-PCR)和红霉素 N-去甲基酶(ERND)测定研究了它们的组织分布以及对镉(Cd)和苯并[a]芘(B[α]P)暴露的反应。 McCYP3A-1 和 McCYP3A-2 具有 CYP 家族的典型结构域,如螺旋-C、螺旋-I、螺旋-K、PERF 和血红素结合结构域,以及 CYP3s 的特征结构域,包括六个 SRS 基序。 McCYP3A-1 和 McCYP3A-2 转录物在所有检测组织中均呈组成型表达,在消化腺、肝胰腺和性腺中表达水平较高。在 B[α]P 暴露下,消化腺中 McCYP3A-1 和 McCYP3A-2 mRNA 表达呈先上调后下调的模式,而在 Cd 暴露下,呈时间依赖性诱导模式。此外,暴露于 Cd 和 B[α]P 后,ERND 活性呈时间依赖性增加。这些结果表明 McCYP3A-1 和 McCYP3A-2 是 CYP3A 家族成员,可能在外源物质代谢清除中发挥潜在作用。同时,目前的结果可为监测多环芳烃和重金属污染提供一些基线数据,支持 McCYP3A-1 和 McCYP3A-2 作为候选生物标志物。

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