Nan Ping, Yan Shuaiguo, Wang Yaxing, Du Qiyan, Chang Zhongjie
Molecular and Genetic Laboratory, College of Life Science, Henan Normal University, No.46, East Jianshe Road, Xinxiang, Henan, 453007, China.
Environ Toxicol. 2017 Feb;32(2):404-416. doi: 10.1002/tox.22244. Epub 2016 Feb 18.
Ionic liquids (ILs) are widely used as reaction media in various commercial applications. Many reports have indicated that most ILs are poorly decomposed by microorganisms and are toxic to aquatic organisms. In this study, differential gene expression profiling was conducted using a suppression subtraction hybridization cDNA library from hepatic tissue of the loach (Paramisgurnus dabryanus) after exposure to 1-hexadecyl-3-methylimidazolium chloride ([C mim]Cl), a representative IL. Two hundred and fifty-nine differentially expressed candidate genes, whose expression was altered by >2.0-fold by the [C mim]Cl treatment, were identified, including 127 upregulated genes and 132 downregulated genes. A gene ontology analysis of the known genes isolated in this study showed that [C mim]Cl-responsive genes were involved in cell cycle, stimulus response, defense response, DNA damage response, oxidative stress responses, and other biological responses. To identify candidate genes that may be involved in [C mim]Cl-induced toxicity, 259 clones were examined by Southern blot macroarray hybridization, and 20 genes were further characterized using quantitative real-time polymerase chain reaction. Finally, six candidate genes were selected, including three DNA damage response genes, two toxic substance metabolic genes, and one stress protein gene. Our results indicate that these changes in gene expression are associated with [C mim]Cl-induced toxicity, and that these six candidate genes can be promising biomarkers for detecting [C mim]Cl-induced toxicity. Therefore, this study demonstrates the use of a powerful assay to identify genes potentially involved in [C mim]Cl toxicity, and it provides a foundation for the further study of related genes and the molecular mechanism of [C mim]Cl toxicity. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 404-416, 2017.
离子液体(ILs)在各种商业应用中被广泛用作反应介质。许多报告表明,大多数离子液体不易被微生物分解,并且对水生生物有毒。在本研究中,利用暴露于代表性离子液体1-十六烷基-3-甲基咪唑氯盐([C mim]Cl)后的泥鳅(Paramisgurnus dabryanus)肝脏组织构建的抑制性消减杂交cDNA文库进行了差异基因表达谱分析。鉴定出259个差异表达的候选基因,其表达因[C mim]Cl处理而改变了2.0倍以上,其中包括127个上调基因和132个下调基因。对本研究中分离出的已知基因进行的基因本体分析表明,[C mim]Cl反应基因参与细胞周期、刺激反应、防御反应、DNA损伤反应、氧化应激反应和其他生物学反应。为了鉴定可能参与[C mim]Cl诱导毒性的候选基因,通过Southern印迹宏阵列杂交检测了259个克隆,并使用定量实时聚合酶链反应进一步表征了20个基因。最后,选择了6个候选基因,包括3个DNA损伤反应基因、2个有毒物质代谢基因和1个应激蛋白基因。我们的结果表明,这些基因表达的变化与[C mim]Cl诱导的毒性有关,并且这6个候选基因有望成为检测[C mim]Cl诱导毒性的生物标志物。因此,本研究展示了一种强大的检测方法来鉴定可能参与[C mim]Cl毒性的基因,并为进一步研究相关基因和[C mim]Cl毒性的分子机制提供了基础。© 2016威利期刊公司。环境毒理学32: 404 - 416, 2017。