Identification of inflammation-related proteins in a murine colitis model by 2D fluorescence difference gel electrophoresis and mass spectrometry
Identification of inflammation-related proteins in a murine colitis model by 2D fluorescence difference gel electrophoresis and mass spectrometry
复制标题
DOI：
10.1111/j.1440-1746.2009.06219.x
复制
发表时间：
2010-05-01
影响因子：
4.1
通讯作者：
Yoshikawa, Toshikazu
中科院分区：
文献类型：
--
作者：
Naito, Yuji;Takagi, Tomohisa;Yoshikawa, Toshikazu
背景和目的：本研究的目的是通过建立小鼠溃疡性结肠炎的体内模型，利用蛋白质组学方法，寻找可能与急性炎症相关的新的肠道蛋白质。方法：采用二维荧光差示凝胶电泳法(2D-DGE)和基质辅助激光解吸电离飞行时间光谱仪(MALDI-TOF)多肽指纹图谱分析正常和炎症小鼠肠黏膜的差异表达蛋白质。用8.0%葡聚糖硫酸钠(DSS)灌胃诱导大鼠急性结肠炎模型。结果：使用吉祥物搜索引擎，在总共7个差异表达的蛋白质点中，我们鉴定了5个不同的蛋白质，其中2个在结肠炎中表达上调，3个在正常粘膜中表达下调。3-羟基-3-甲基戊二酰辅酶A合成酶2和丝氨酸B1a表达上调，蛋白二硫键异构酶A3、过氧化还蛋白-6和波形蛋白表达下调。结论：这些蛋白可能与肠炎的发生有关。2D-DGE和MALDI-TOF质谱学在寻找差异表达蛋白质方面是有用的。
Background and Aims:The aim of this study was to identify new intestinal proteins potentially associated with acute inflammation using proteomic profiling of an in vivo mice model of ulcerative colitis.Methods:2D fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time-of-flight spectrometer (MALDI-TOF) peptide mass fingerprinting were used to determine differentially expressed proteins between normal and inflamed intestinal mucosa. Acute colitis was induced by 8.0% dextran sodium sulfate (DSS) given p.o. for 7 days.Results:Among a total of seven protein spots showing differential expression, we identified five different proteins, of which two were upregulated and three downregulated in colitis in comparison to normal mucosa, using the MASCOT search engine. 3-Hydroxy-3-methylglutaryl-coenzyme A synthase 2 and serpin b1a were upregulated proteins, and protein disulfide-isomerase A3, peroxiredoxin-6 and vimentin were identified as downregulated proteins.Conclusion:These identified proteins may be responsible for the development of the intestinal inflammation. 2D-DIGE and MALDI-TOF mass spectrometry are useful in the search for the differentially expressed proteins.