同位素标记相对和绝对定量技术在RA患者血清中的应用研究

Application of iTRAQ technology on analysis of serum from the patients with RA

采用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantitation, iTRAQ)技术寻找能够用于RA诊断、疗效评估或预后判断的新血清标志物,探索RA发病机制。对10例活动期RA患者、10例稳定期RA患者与10例健康对照者血清采用iTRAQ技术结合串联质谱技术进行差异蛋白质组学分析,得到差异蛋白。通过DAVID软件鉴定差异蛋白,分析差异蛋白在RA发病过程中所起的生物学作用。结果显示共鉴定出303个蛋白质,其中RA活动组与健康对照组差异倍数≥1.5的蛋白质27个,表达显著上调和下调的蛋白质分别为13和14个,差异蛋白的功能涉及到蛋白质代谢、脂类代谢、补体系统、凝血系统等多个系统。通过DAVID软件的在线分析得到了差异蛋白的基因本体论(gene ontology, GO)生物学过程、细胞组分和分子功能的富集分析结果。通过对差异蛋白的研究可以为阐明RA的发病机制及发现新的生物标志物提供新的途径。

The quantitative proteomics technology based on the isobaric tags for relative and absolute quantitation(iTRAQ) was used to find new serum markers for RA diagnosis, evaluation of treatment effect or prognosis and explore the mechanism of pathogenesis. Twenty patients with RA, 10 in active stage and 10 in stable stage, were enrolled. In addition, 10 healthy individuals were as controls. Sera were collected from each individual and analyzed by the iTRAQ technology and tandem mass spectrometry to get differential proteins. Such differential proteins were identified and their biological roles in the pathogenesis of RA were analyzed by the DAVID software. A total of 303 proteins were identified and analyzed. The difference between the active group and the control group was more than or equal to 1.5 folds in 27 proteins, of which, 13 proteins upregulated and 14 downregulated. The differential proteins were involved in protein metabolism, lipid metabolism, complement system and blood coagulation system. With online analysis of DAVID software, the enrichment analysis results of the biological process of the gene ontology(GO), cellular component and molecular function for the differential protein were obtained. Studying of these differential proteins can provide a new means to understand the pathogenesis of RA and to discover new biomarkers.