Department of Biotechnology and Life Sciences, University of Insubria, I-21100 Varese, Italy.
Toxicol Lett. 2013 Apr 12;218(2):118-28. doi: 10.1016/j.toxlet.2013.01.023. Epub 2013 Feb 7.
Cigarette smoke is a significant independent risk factor for vascular diseases and is a leading cause of structural and functional alterations of the vascular endothelium. In this study, we show protein carbonylation in the human umbilical vein endothelial cell line (ECV-304) exposed to whole-phase cigarette smoke extract. The main carbonylated proteins, including cytoskeletal proteins, glycolytic enzymes, xenobiotic metabolizing and antioxidant enzymes, and endoplasmic reticulum proteins, were identified by means of two-dimensional electrophoresis and Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) mass spectrometry (redox proteomics). Morphological analyses by fluorescence microscopy evidenced alterations in the microtubule cytoskeleton, especially at longer exposure time to cigarette smoke extract. Morphological analyses by transmission electron microscopy showed vacuolisation of the cytoplasm, alteration of mitochondria ultrastructure, and some enlargement of the perinuclear space. The possible role played by protein carbonylation caused by reactive species contained in cigarette smoke in the cigarette smoke-induced endothelial injury is discussed.
香烟烟雾是血管疾病的一个重要独立风险因素，也是血管内皮结构和功能改变的主要原因。在这项研究中，我们展示了暴露于全相香烟烟雾提取物的人脐静脉内皮细胞系（ECV-304）中的蛋白质羰基化。通过二维电泳和基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱（氧化还原蛋白质组学）鉴定了主要的羰基化蛋白质，包括细胞骨架蛋白、糖酵解酶、外源性代谢和抗氧化酶以及内质网蛋白。荧光显微镜的形态分析表明微管细胞骨架发生了改变，尤其是在更长时间暴露于香烟烟雾提取物后。透射电子显微镜的形态分析显示细胞质空泡化、线粒体超微结构改变以及核周空间有些扩大。讨论了香烟烟雾中活性物质引起的蛋白质羰基化在香烟烟雾诱导的内皮损伤中可能发挥的作用。
