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通过 2D 荧光差异凝胶电泳和质谱鉴定小鼠结肠炎模型中的炎症相关蛋白。

Identification of inflammation-related proteins in a murine colitis model by 2D fluorescence difference gel electrophoresis and mass spectrometry.

机构信息

Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto, Japan.

出版信息

J Gastroenterol Hepatol. 2010 May;25 Suppl 1:S144-8. doi: 10.1111/j.1440-1746.2009.06219.x.

Abstract

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.

摘要

背景与目的

本研究旨在通过溃疡性结肠炎动物模型的蛋白质组学分析,鉴定与急性炎症相关的新肠道蛋白。

方法

采用二维荧光差异凝胶电泳(2D-DIGE)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)肽质量指纹图谱技术,检测正常和炎症性肠黏膜之间差异表达的蛋白质。采用口服 8.0%葡聚糖硫酸钠(DSS)诱导急性结肠炎,连续 7 天。

结果

在总共 7 个显示差异表达的蛋白斑点中,我们使用 MASCOT 搜索引擎鉴定了 5 种不同的蛋白质,其中 2 种在结肠炎中上调,3 种在结肠炎中下调,与正常黏膜相比。3-羟-3-甲基戊二酰辅酶 A 合酶 2 和丝氨酸蛋白酶抑制剂 b1a 为上调蛋白,而蛋白二硫键异构酶 A3、过氧化物酶 6 和波形蛋白为下调蛋白。

结论

这些鉴定的蛋白可能与肠道炎症的发生有关。2D-DIGE 和 MALDI-TOF 质谱分析在寻找差异表达蛋白方面非常有用。

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