S100A4, A10在类风湿关节炎滑膜成纤维细胞中表达及意义.docVIP

S100A4, A10在类风湿关节炎滑膜成纤维细胞中表达及意义 　 作者：柏干苹，周丽娜，贺伟峰，罗高兴，陈烯伟，陈光兴，胡晓红，石东文，方勇飞，吴军 【摘要】 目的：探讨正常人与类风湿关节炎(RA)患者滑膜成纤维细胞蛋白质组表达差异，分析差异蛋白在RA发病中的作用. 方法：应用固相pH梯度(IPG)双向凝胶电泳(2DE)分离正常人及RA患者滑膜成纤维细胞总蛋白质；应用双向电泳凝胶分析软件（PDQuest 7.3.1）对2DE凝胶进行量化比较分析；应用基质辅助激光解析电离飞行时间质谱鉴定部分差异蛋白质，并用Western Blot、免疫细胞化学技术验证表达差异蛋白质. 结果：正常人及RA患者滑膜成纤维细胞蛋白2DE凝胶图谱上分别平均显示896和992个点. 有64个蛋白质点表达存在2倍以上的差异. 选取9个在RA滑膜成纤维细胞中表达上调的蛋白质点进行质谱鉴定，成功鉴定出钙结合蛋白S100A4和S100A10两个蛋白质. 经Western Blot、免疫细胞化学技术验证这两个蛋白在RA滑膜成纤维细胞中表达上调，与上述结果一致，并且发现S100A4在RA滑膜成纤维细胞中的表达分布发生了变化. 结论：结合蛋白S100A4和S100A10可能参与RA的发病. S100A4在细胞中的表达易位可能与RA滑膜成纤维细胞表型改变有关. 【关键词】 滑膜成纤维细胞；蛋白质组；双向电泳；类风湿关节炎；S100A4基因；S100A10基因 　　【Abstract】 AIM: To find the differentially expressed proteins of synovial fibroblasts (SFs) from rheumatoid arthritis (RA) patients and normal controls using twodimensional gel electrophoresis (2DE) and bioinformatics for providing useful and new clues to elucidate the pathogenesis and study the treatment of RA. METHODS: The total protein of SFs from either RA patients or normal ones was prepared by means of immobilized pH gradient based on 2DE. Gels were scanned, and the images were processed with PDQuest 7.3.1 software. The differentiallly expressed proteins were identified by matrixassisted laser desorption ionizationtimeofflight mass spectrometry (MALDITOFMS). Then some of them were validated by Western Blot and immunocytochemistry. RESULTS: 896 protein spots were detected in SFs of normal individuals, and 992 spots in RA, respectively. There were 64 different protein spots. A total of 9 protein spots that expressed at higher levels in SFs from RA were identified by MALDITOFMS, and 2 valid proteins were obtained. They were calcium binding protein S100A4 and S100A10. Western Blot and immunocytochemistry were used to further verify differentially expressed proteins. The results showed that expression levels of S100A4 and S100A10 were significantly higher in SFs from RA than in SFs from normal individuals. Furthermore, the distribution of S1