Disorders in Angiogenesis and Redox Pathways Are Main Factors Contributing to the Progression of Rheumatoid Arthritis A Comparative Proteomics Study
Disorders in Angiogenesis and Redox Pathways Are Main Factors Contributing to the Progression of Rheumatoid Arthritis A Comparative Proteomics Study
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DOI：
10.1002/art.33425
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发表时间：
2012-04-01
影响因子：
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通讯作者：
Jiao, Bing-Hua
中科院分区：
文献类型：
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作者：
Wang, Jian-Guang;Xu, Wei-Dong;Jiao, Bing-Hua
目标。采用蛋白质组学分析方法，比较类风湿关节炎（RA）患者成纤维细胞样滑膜细胞（FLS）与正常人FLS的蛋白表达，探讨类风湿关节炎（RA）的发病机制。采用自动二维纳米电喷雾液相色谱串联质谱法对50例RA患者和10例正常人的FLS原代培养物中提取的蛋白质进行了分析。基于生物信息学分析，采用2样本t检验（P < 0.05）和fold change（> 1.5）筛选差异表达蛋白。实时聚合酶链反应和Western blotting检测滑膜组织中血管发育相关蛋白(Thy-1、结缔组织生长因子（CTGF）、血栓反应蛋白1 （TSP-1）和氧化还原相关蛋白（超氧化物歧化酶2 (SOD2)）的表达。通过RNA干扰实验分析Thy-1和CTGF下调对Thy-1、CTGF、TSP-1和血管内皮生长因子（VEGF）的影响。根据每个蛋白质存在bb0.1个独特肽和错误发现率为的标准
Objective. To clarify the pathogenesis of rheumatoid arthritis (RA) by comparing protein expression in fibroblast-like synoviocytes (FLS) from patients with RA with that in FLS from normal subjects, using proteomics analysis.Methods. Proteins extracted from primary cultures of FLS obtained from 50 patients with RA and 10 normal subjects were analyzed by automated 2-dimensional nano-electrospray ionization liquid chromatography tandem mass spectometry. Differentially expressed proteins were screened by 2-sample t-test (P < 0.05) and fold change (> 1.5), based on the bioinformatics analysis. The expression of vasculature development-related proteins (Thy-1, connective tissue growth factor [CTGF], and thrombospondin 1 [TSP-1]) and redox-related proteins (superoxide dismutase 2 [SOD2]) in synovial tissue was confirmed by real-time polymerase chain reaction and Western blotting. The effect of Thy-1 and CTGF knockdown on Thy-1, CTGF, TSP-1, and vascular endothelial growth factor (VEGF) was analyzed by RNA interference experiments.Results. According to the criteria of having > 1 unique peptide per protein present and a false discovery rate of