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在新的虎鱼(短吻背眼虾虎鱼)海洋鱼类模型中,苯并[a]芘胁迫介导的转录反应特征分析

Characterization of transcriptional responses mediated by benzo[a]pyrene stress in a new marine fish model of goby, Mugilogobius chulae.

作者信息

Cai Lei, Li Jianjun, Yu Lujun, Wei Yuanzheng, Miao Zongyu, Chen Meili, Huang Ren

机构信息

Key Laboratory of Guangdong Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, No. 11, Fengxin Road, Huangpu District, Guangzhou, 510663, Guangdong, People's Republic of China.

出版信息

Genes Genomics. 2019 Jan;41(1):113-123. doi: 10.1007/s13258-018-0743-8. Epub 2018 Sep 21.

Abstract

Benzo[a]pyrene (BaP) is one of the most studied targets among polycyclic aromatic hydrocarbons (PAHs). Because of the complexity of the toxicity mechanism in BaP, little is known about the molecular mechanism at the level of transcription of BaP in marine fishes. The primary objective of this study was to investigate the molecular basis of the effects of BaP on marine fish, using Mugilogobius chulae (Smith 1932) as the model. A closed colony of M. chulae was used for the BaP toxicity test. Two fish liver samples per replicate from each group were excised and blended into one sample by pooling an equal amount of liver tissue. Total RNA of all samples was extracted separately. Equal quantities of total RNA from the three replicates of the two groups were pooled for sequencing. The sequencing cDNA libraries were sequenced using Illumina HiSeq 2000 system. Differentially expressed genes were detected with the DEGSeq R package. In total, 52,364,032 and 53,771,748 clean nucleotide reads were obtained in the control and BaP-exposed libraries, respectively, with N50 lengths of 1277 and 1288 bp, respectively. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed a significant enrichment of genes related to detoxification, transportation, and lipid metabolism. We also identified, for the first time, an association between endoplasmic reticulum dysfunction and lipid metabolism resulting from BaP exposure. Using quantitative real-time PCR, some effective molecular biomarkers for monitoring of BaP-polluted seawater were identified. The results demonstrate that BaP enhanced the expression of genes involved in detoxification in M. chulae and inhibited that of genes related to lipid metabolism, possibly by suppressing the expression of numerous ER-related genes involved in fat digestion and absorption.

摘要

苯并[a]芘(BaP)是多环芳烃(PAHs)中研究最多的目标之一。由于BaP毒性机制的复杂性,关于其在海洋鱼类转录水平上的分子机制知之甚少。本研究的主要目的是使用中华间吸鳅(Smith,1932年)作为模型,研究BaP对海洋鱼类影响的分子基础。使用中华间吸鳅的一个封闭群体进行BaP毒性试验。每组每次重复取两个鱼肝样本,将等量的肝组织汇集到一个样本中并混合。分别提取所有样本的总RNA。将两组三个重复的等量总RNA汇集用于测序。使用Illumina HiSeq 2000系统对测序cDNA文库进行测序。使用DEGSeq R包检测差异表达基因。在对照组和暴露于BaP的文库中,分别获得了52364032和53771748条干净的核苷酸读数,N50长度分别为1277和1288 bp。基因本体论和京都基因与基因组百科全书通路分析显示,与解毒、运输和脂质代谢相关的基因显著富集。我们还首次确定了内质网功能障碍与BaP暴露导致的脂质代谢之间的关联。使用定量实时PCR,鉴定了一些用于监测BaP污染海水的有效分子生物标志物。结果表明,BaP增强了中华间吸鳅中参与解毒的基因的表达,并抑制了与脂质代谢相关的基因的表达,可能是通过抑制许多参与脂肪消化和吸收的内质网相关基因的表达来实现的。

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