S100A4,A10在类风湿关节炎滑膜成纤维细胞中的表达和意义.docVIP

S100A4, A10在类风湿关节炎滑膜成纤维细胞中的表达及意义 　　作者：柏干苹,周丽娜,贺伟峰,罗高兴,陈烯伟　　作者单位：第三军医大学西南医院：烧伤研究所, 关节外科中心，风湿科，重庆 400038 　　【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 　　Expressions and significance of S100A4 and S100A10 in synovial fibroblasts obtained from rheumatoid arthritis patients 　　BO GanPing, ZHOU LiNa, HE WeiFeng, LUO GaoXing, CHEN XiWei, CHEN GuangXing, HU XiaoHong, SHI DongWen, FANG YongFei, WU Jun 　　Institute of Burn Research, Department of Orthopaedics, Department of Rheumatism, Southwest Hospital, Third Military Medical University, Chongqing 400038, China 　　【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