类风湿性关节炎患者脑脊液中疾病相关蛋白的二维电泳分析

Yukio Yamamoto, Y. Akita, Shigeyuki Tai, S. Fukasaku, Teruhide Yamaguchi, T. Oshizawa, K. Yamaoka, M. Shimamura, T. Hazato
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引用次数: 2

摘要

比较类风湿关节炎(RA)患者与对照组脑脊液(CSF)中的蛋白质表达,可以揭示影响疾病进展和调节对药物反应的蛋白质。二维凝胶电泳(2-DE)和银染色用于鉴定RA患者疾病相关的CSF蛋白。首先,为了加强对脑脊液蛋白的检测,并提高2-DE对其亚型的分离,脑脊液样品先用白蛋白和IgG去除试剂盒预处理,然后用丙酮沉淀。2-DE分析显示脑脊液中白蛋白和免疫球蛋白有1600多个斑点。两组蛋白斑点的表达均无明显变化,但两组RA样品中蛋白斑点的表达均有明显变化。特别是,在RA样品中,大约50 kD的蛋白表达显著增加,而10-15 kD和中性pI的两个序列蛋白点的表达减少。这些初步结果表明,蛋白质组学方法有助于阐明RA危机的机制,一些表达改变的蛋白可能是RA疾病相关蛋白的新候选蛋白。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Two-dimensional electrophoretic analysis of disease-associated proteins in human cerebrospinal fluid from patients with rheumatoid arthritis
Comparing protein expression in the cerebrospinal fluid (CSF) of rheumatoid arthritis (RA) patients with that of controls, makes possible the uncovering of proteins that affect disease progression and regulate responsiveness to drugs. Two-dimensional gel electrophoresis (2-DE) and silver staining were used for identifying disease-associated CSF proteins in RA patients. First, to enhance the detection of CSF proteins and to improve the separation of their isoforms by 2-DE, CSF samples were pre-treated with an albumin and IgG removal kit, then by acetone precipitation. The 2-DE analysis revealed more than 1600 spots by the removal of albumin and immunoglobulin from CSF. The expression of the protein spots was not greatly changed in either group, but some notable changes in protein spots were observed in two RA samples. In particular, the expression of an approximately 50 kD protein increased markedly, whereas that of two sequential protein spots of 10-15 kD and with neutral pI decreased in the RA samples. These preliminary results suggest that the proteomic method is conducive to clarifying the mechanism of RA crises, and that some of the expression-changed proteins may be new candidates for disease-associated proteins of RA.
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