双向电泳和质谱技术在类风湿关节炎滑膜细胞差异蛋白分析中的应用及意义

Screening differentially expressed proteins of fibroblast-like synoviocytes in rheumatoid arthritis by 2-DE and MALDI-TOF-MS

目的 采用2-DE和质谱技术分析RA患者和创伤件关节炎患者关节滑膜组织中FLS蛋白质组学的变化,寻找RA关节滑膜特异性自身抗原.方法 采用2-DE分离6例创伤性关节炎患者及3例RA患者关节滑膜组织中FLS总蛋白质,凝胶经银染显色和图像数据分析识别差异蛋白质点,并用MALDI-TOF-MS对筛选出的差异蛋白质点进行鉴定.结果 RA患者与创伤性关节炎患者中的FLS蛋白质通过2-DE检测,分别检出1 633、1 603个蛋白质点;对2组相互匹配的蛋白质点进行差异分析后,其中蛋白质差异量大于3倍的差异点共有92个,从中选取在RA患者FLS中表达上调的33个差异蛋白质点进行MALDI-TOF-MS 鉴定分析,并与IPI和Swiss-Prot数据库进行比对,共鉴定出包括肿瘤型丙酮酸激酶、α-烯醇化酶、二硫键异构酶A3 前体、谷胱甘肽转移酶、泛素结合酶E2K、血小板活化因子乙酰水解酶、二氢嘧啶酶样2、腺苷酸酶、转醛醇酶、乙酰基辅酶A异构酶、26S蛋白调节亚单位S6a、膜联蛋白A11、过氧化物还原酶1、人色氨酰tRNA合成酶、核纤层蛋白、肌间蛋白、核基质蛋白、肌动蛋白、T-复合物多肽1、葡萄糖调节蛋白75、αB-晶体蛋白、葡萄糖调节蛋白78、未知蛋白：如PSMFA和SELENBP1等30个与RA发病有关的蛋白.结论 RA患者与创伤性关节炎患者FLS间存在差异表达的蛋白质,这些差异蛋白有可能是构成RA关节滑膜的相关自身抗原,并可能在诱导RA自身免疫反应中发挥一定作用.

Objective To identify synovium-associated proteins of by analyzing proteome of FLS between RA patients and traumatic arthritis. Methods The total protein of FLS from 3 patients of RA and 6 normal controls were separated by 2-DE. The differentially expressed proteins were identified by MALDI-TOF-MS. Results The 2-DE image results showed that there were many proteins differentially expressed in FLS from RA and control. A total of 33 differential proteins that were up-regulated in RA as compared with controls were selected and 30 proteins including pyruvate kinase isozymes M1/M2, α-enolase, protein disulfide-isomerase A3 precursor, glutathione transferase, peroxiredoxin 1, ubiquitin-conjugating enzyme E2K, platelet-activating factor acetylhydrolase IB, dihydropyrimidinase-related protein 2, adenosine kinase, transaldolase, δ-δ-dienoyl-CoAisomerase, 26S protease regulatory subunit 6A, annexin All, tryptophanyl-tRNA synthetase isoform b, lamin-A/C, myomesin-1 isoform b, ruvB-like 1, actin-cytoplasmic 1, T-complex protein 1, stress-70 protein, mitochondrial precursor, α-crystallin B chain, 78 000 glucose-regulated protein, PSME4, SELENBP1 and so on. Conclusion The differentially expressed proteins are present in FLS from RA and control, which may be the synovium biomakers and involved in pathogenesis of RA.