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甲基对硫磷暴露于斑马鱼肝细胞中蛋白质图谱的改变:一种膜蛋白质组学方法。

Alterations of protein profile in zebrafish liver cells exposed to methyl parathion: a membrane proteomics approach.

机构信息

State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen 361005, China.

出版信息

Chemosphere. 2012 Mar;87(1):68-76. doi: 10.1016/j.chemosphere.2011.11.061. Epub 2011 Dec 17.

Abstract

Methyl parathion (MP) is an extensively used organophosphorus pesticide, which has been associated with a wide spectrum of toxic effects on environmental organisms. The aim of this study is to investigate the alterations of membrane protein profiles in zebrafish liver (ZFL) cell line exposed to MP for 24 h using proteomic approaches. Two-dimensional gel electrophoresis revealed a total of 13 protein spots, whose expression levels were significantly altered by MP. These differential proteins were subjected to matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry analysis, and nine proteins were identified to be membrane proteins, among which seven were up-regulated, while two were down-regulated. In addition, the mRNA levels corresponding to these differential membrane proteins were further analyzed by quantitative real-time PCR. And the differential expression of arginase-2 was specially validated via Western blotting. Regarding the physiological functions, these proteins are involved in molecular chaperon, cytoskeleton system, cell metabolism, signal transduction, transport and hormone receptor respectively, suggesting the complexity of MP-mediated toxicity to ZFL cell. These data could provide useful insights for better understanding the hepatotoxic mechanisms of MP and develop novel protein biomarkers for effectively monitoring MP contamination level in aquatic environment.

摘要

甲基对硫磷(MP)是一种广泛使用的有机磷农药,它对环境生物具有广泛的毒性作用。本研究旨在采用蛋白质组学方法研究甲基对硫磷暴露 24 小时后斑马鱼肝脏(ZFL)细胞系中膜蛋白谱的变化。二维凝胶电泳共显示出 13 个蛋白斑点,其表达水平受 MP 显著改变。这些差异蛋白经基质辅助激光解吸/电离串联飞行时间质谱分析鉴定为膜蛋白,其中 7 个上调,2 个下调。此外,通过定量实时 PCR 进一步分析了这些差异膜蛋白对应的 mRNA 水平。通过 Western 印迹专门验证了精氨酸酶-2 的差异表达。就生理功能而言,这些蛋白分别涉及分子伴侣、细胞骨架系统、细胞代谢、信号转导、运输和激素受体,表明 MP 对 ZFL 细胞的毒性作用的复杂性。这些数据可以为更好地理解 MP 的肝毒性机制提供有用的见解,并为有效监测水环境中 MP 的污染水平开发新的蛋白质生物标志物。

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