类风湿性关节炎患者关节滑膜组织蛋白质组学

背景：已有研究表明滑膜炎持久反复发作,最终导致关节内软骨和骨的破坏,国内关于类风湿性关节炎滑膜组织的蛋白质组学研究报道较少。 目的：通过比较类风湿性关节炎患者和无关节滑膜损伤患者滑膜组织蛋白质表达的差异,探讨类风湿性关节炎可能的发病机制,寻找类风湿性关节炎致病相关蛋白。 方法：选取6例无关节滑膜损伤患者及6例活动期类风湿性关节炎患者行关节镜手术得到的滑膜组织,提取总蛋白质后进行双向凝胶电泳,考马斯亮蓝染色,PDQUEST软件分析,对差异蛋白质点采用基质辅助激光解析电离质谱(MALDI-TOF-MS)技术进行鉴定,并用Mascot软件在SwissProt和NCBInr数据库中进行同源比较和分析鉴定。 结果与结论：建立了类风湿性关节炎组及对照组滑膜组织双向凝胶电泳图谱,获得130个差异在2倍以上的蛋白质点数,选取其中分辨清楚的39个点进行鉴定,初步鉴定出29个蛋白质,其中21个蛋白质点在类风湿性关节炎组中表达上调,8个表达下调,其功能涉及功能代谢、细胞信号传导、抗氧化、分子伴侣等。结果表明类风湿关节炎滑膜病变是一个多种蛋白质参与的复杂过程,这些表达差异蛋白质可能是类风湿性关节炎发病的内在因素。

Proteomics research on the joint synovium of patients with rheumatoid arthritis

BACKGROUND:Several studies have shown that it can result in joint cartilages and bones breaking due to the sustained attack of synovitis. But reports about proteomics research on the joint synovium of rheumatoid arthritis (RA) patients in China are few. OBJECTIVE:To explore the possible underlying mechanism of RA and to seek out RA associated proteins by comparing protein expression difference between the synovium of patients with surgical trauma and patients with RA. METHODS:The synovial tissues of six patients with surgical trauma and six patients with RA were obtained by arthroscopic surgery and they were divided into RA group and control group. The total proteins were extracted from synovial tissue and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE), which was followed by Coomassie Brilliant Blue staining to visualize the protein spots in the 2-DE. The differentially expressed proteins were analyzed by using PDQUEST analysis software, and then identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and homogonously compared and identified in Swiss Prot and NCBInr databases with Mascot software. RESULTS AND CONCLUSION:The 2-DE patterns of RA group and control group were established. Proteins were separated by the 2-DE, and 130 proteins were found differentially expressed over two folds between synovium of surgical trauma and RA patients. Thirty-nine clear proteins of them were selected and analyzed with MALDI-TOF-MS and 29 proteins were identified. Among the identified differential expression proteins, 21 proteins were up-regulated and eight were down-regulated, involved in substance metabolism, signaling pathway, anti-oxidation protection and molecule chaperone. These findings suggest that synovium lesions of RA are a complex process in which multiple proteins are involved, and some differentially expressed proteins may play a potential role in the pathogenesis of RA.