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寻找在体内接触全氟辛烷磺酸和真实世界野外研究后,欧鳗外周血单核细胞中的蛋白质表达特征。

Looking for protein expression signatures in European eel peripheral blood mononuclear cells after in vivo exposure to perfluorooctane sulfonate and a real world field study.

机构信息

Research Unit in Environmental and Evolutionary Biology (URBE), NARILIS (Namur Research Institute for Life Sciences), University of Namur, Belgium.

出版信息

Sci Total Environ. 2014 Jan 15;468-469:958-67. doi: 10.1016/j.scitotenv.2013.07.110. Epub 2013 Oct 2.

Abstract

The decline of European eel population can be attributed to many factors such as pollution by xenobiotics present in domestic and industrial effluents. Perfluorooctane sulfonate (PFOS) is a ubiquitous compound of a particular concern in Europe. PFOS can reach high concentrations in tissues of organisms and many toxic effects have been reported in fish. This study aimed at evaluating the toxicological effects of PFOS in European eel peripheral blood mononuclear cells (PBMCs) at the protein expression level. To identify proteins whose expression was modified by PFOS, we performed a proteomic analysis on the post-nuclear fraction of PBMCs after a chronic exposure (28 days) of yellow eels to zero, 1 or 10 μg/L PFOS. This in vivo study was completed by a proteomic field study on eels sampled in Belgian rivers presenting different PFOS pollution degrees. Proteins were separated by two-dimensional in-gel electrophoresis (2D-DIGE) to compare the post-nuclear fraction of PBMCs from the reference group with cells from fish exposed to the pollutant of interest. On the 28 spots that were significantly (p < 0.05; ANOVA followed by a Dunnett post-hoc test) affected by PFOS in the in vivo experiment, a total of 17 different proteins were identified using nano-LC ESI-MS/MS and the Peptide and Protein Prophet of Scaffold software. In the field experiment, 18 significantly (p < 0.05; ANOVA followed by Dunnett's test) affected spots conducted to the identification of 16 different proteins. Interestingly, only three proteins were found in common between in vivo and in situ experiments: plastin-2, alpha-enolase and glyceraldehyde 3-phosphate dehydrogenase. Comparing the results with a previous study, plastin-2 and alpha-enolase were also been found to be affected after in vitro exposure of PBMCs during 48 h to either 10 μg or 1 mg PFOS/L. Potential use of these proteins as biomarkers of PFOS exposure in European eel could indicate early warning signals.

摘要

欧洲鳗种群的减少可归因于许多因素,如生活污水和工业废水中存在的异生物质污染。全氟辛烷磺酸 (PFOS) 是一种在欧洲特别受关注的普遍存在的化合物。PFOS 可以在生物体组织中达到很高的浓度,并在鱼类中报告了许多毒性作用。本研究旨在评估 PFOS 在欧洲鳗外周血单核细胞 (PBMC) 中的蛋白质表达水平上的毒理学效应。为了确定 PFOS 改变表达的蛋白质,我们对慢性暴露(28 天)于零、1 或 10μg/L PFOS 的黄鳗 PBMC 的核后部分进行了蛋白质组分析。这项体内研究通过对在比利时河流中采样的鳗鱼进行蛋白质组学野外研究得到了补充,这些河流的 PFOS 污染程度不同。通过二维凝胶电泳(2D-DIGE)分离蛋白质,以比较参考组 PBMC 的核后部分与暴露于污染物的鱼类细胞。在体内实验中,有 28 个斑点受到 PFOS 的显著影响(p < 0.05;ANOVA 后 Dunnett 事后检验),使用纳升 LC ESI-MS/MS 和 Peptide and Protein Prophet of Scaffold 软件共鉴定出 17 种不同的蛋白质。在野外实验中,18 个显著(p < 0.05;ANOVA 后 Dunnett 检验)受影响的斑点鉴定出 16 种不同的蛋白质。有趣的是,只有三种蛋白质在体内和原位实验中是共同的:韧丝蛋白-2、α-烯醇酶和甘油醛 3-磷酸脱氢酶。将结果与之前的一项研究进行比较,在体外暴露于 PBMC 48 小时后,无论是 10μg 还是 1mg PFOS/L,韧丝蛋白-2 和α-烯醇酶也被发现受到影响。这些蛋白质可能作为欧洲鳗 PFOS 暴露的生物标志物,可指示早期预警信号。

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