质谱技术对类风湿关节炎患者滑膜免疫复合物差异蛋白质的分析

Differential proteomic analysis for immune complexes deposited in synovial tissues of RA patients using liquid chromatography-tandem mass spectrometry

目的利用质谱技术对类风湿关节炎(RA)患者关节滑膜中的免疫复合物进行全面的抗原谱分析。方法选取本院3例RA患者和3例骨关节炎(osteoarthritis,OA)患者的关节滑膜,通过蛋白质提取、免疫沉淀、胶内酶解和液相色谱串联质谱系统,对滑膜免疫复合物进行抗原谱分析。用FOT值(fraction of total)评价蛋白质的丰度,通过FOT值的比较筛选出上调蛋白和下调蛋白。用David和String软件对差异蛋白质进行功能富集、相互作用网络和信号通路分析。结果在RA和OA患者的关节滑膜组织免疫复合物中分别鉴定出511和526种蛋白质。通过维恩图比较,仅在RA组出现的蛋白质有170种。RA组有45种蛋白质发生上调,85种蛋白质发生下调。结论热休克蛋白HSP90AA1、HSP70、人类白细胞抗原(HLA)-G、硫氧还蛋白、膜联蛋白A2和玻连蛋白可能通过不同的机制参与RA的发病过程,具有RA新的生物标志物的潜在应用价值。

Objective To scan protein expression profile of immune complexes（ ICs） derived from the synovial tissue of the patients with rheumatoid arthritis（ RA） based on liquid chromatography-tandem mass spectrometry（ LC-MS）. Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis（ OA） used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University. The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis. The value of fraction of total（ FOT） was used to estimate protein abundance and screen the up-and down-regulated proteins. The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String. Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively. Among them,170 proteins existed only in RA group. 45 and85 proteins in RA group were statistically up-and down-expressed compared with controls. Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.