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采用抑制性消减杂交和新一代测序技术研究三氯生和混合三氯生及甲基三氯生对斑马鱼胚胎的生态毒基因分析。

Ecotoxicogenomic analysis of zebrafish embryos exposed to triclosan and mixture triclosan and methyl triclosan using suppression subtractive hybridization and next-generation sequencing.

机构信息

Department of Civil and Environmental Engineering, National University of Singapore, Singapore.

Department of Biological Sciences, National University of Singapore, Singapore.

出版信息

J Hazard Mater. 2021 Jul 15;414:125450. doi: 10.1016/j.jhazmat.2021.125450. Epub 2021 Feb 26.

Abstract

Triclosan (TCS) and methyl-triclosan (MTCS), an environmental transformation product of biocide of TCS, have been detected in water, sediment, fish, and invertebrates. In this study, the key pathway perturbation in zebrafish (Danio rerio) embryos exposed to TCS (300 μg/L) and TCS/MTCS mixture (300 μg/L TCS + 30 μg/L MTCS) was assessed by integrating the metabolomic and transcriptomic dysregulation. The differential expressed genes (DEGs) were obtained from the subtracted cDNA libraries by using the suppression subtractive hybridization and next-generation sequencing approach. The dysregulation of twenty-eight GO terms and four KEGG pathways, including oxidative phosphorylation and cardiac muscle contraction, were shown in the TCS treatment group, indicating that TCS could disrupt the mitochondrial inner membrane function by downshifting the electrochemical gradient. Meanwhile, the addition of MTCS in the exposure would cause fourteen additional significant KEGG pathway changes, demonstrating the different effects between two exposure. A pathway-based analysis using the identified DEGs and the altered metabolites in zebrafish embryos treated with TCS and TCS/MTCS mixture, collectively, has been applied. This study demonstrated that the integration of SSH-NGS and metabolomics could reveal toxic effects and potential diseases associated with the exposures of TCS and MTCS in aquatic environments.

摘要

三氯生(TCS)和甲基三氯生(MTCS)是一种生物杀灭剂 TCS 的环境转化产物,已在水、沉积物、鱼类和无脊椎动物中被检测到。在这项研究中,通过整合代谢组学和转录组学失调,评估了斑马鱼(Danio rerio)胚胎暴露于 TCS(300μg/L)和 TCS/MTCS 混合物(300μg/L TCS+30μg/L MTCS)中的关键途径扰动。通过抑制性消减杂交和下一代测序方法,从消减 cDNA 文库中获得差异表达基因(DEGs)。在 TCS 处理组中,观察到 28 个 GO 术语和 4 个 KEGG 途径的失调,包括氧化磷酸化和心肌收缩,表明 TCS 可以通过降低电化学梯度来破坏线粒体内膜功能。同时,在暴露中添加 MTCS 会导致另外 14 个显著的 KEGG 途径变化,表明两种暴露的影响不同。本研究采用基于途径的分析方法,利用鉴定的 DEGs 和 TCS 和 TCS/MTCS 混合物处理的斑马鱼胚胎中的代谢物变化,进行了综合分析。这项研究表明,SSH-NGS 和代谢组学的整合可以揭示 TCS 和 MTCS 在水生环境中的暴露所带来的毒性作用和潜在疾病。

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