BACKGROUND:There is lack of effective diagnostic methods for early knee osteoarthritis. Proteomics refers to the large-scale experimental analysis of proteomes from the overal level of intracel ular protein compositioon, expression and modification, based on which, we can understand protein-protein interaction and relationship, thereby revealing protein functions and cel activity patterns. OBJECTIVE:To apply the isobaric tags for relative and absolute quantitation (iTRAQ) to identify proteins differential y in serums of osteoarthritis Kel gren-Lawrence classification at each stage, and to find out potential molecular markers at each stage of osteoarthritis. METHODS:Sixty patients with knee osteoarthritis were enrol ed, and according to Kel gren-Lawrence (K-L) grading, they were subdivided into K-L 0, II and IV groups. There were 10 males and 10 females randomly selected in each group. Proteins differential y regulated in serums were identified by the stable isotopes 116, 117, 118 of the iTRAQ labeled, reversed-phase column separation, mass spectrometry and Swissport database search, after the serums were subjected to high abundance proteins depletion. Final y, we analyzed the proteins identified by using bioinformatics software. RESULTS AND CONCLUSION:Total y 169 proteins were identified through iTRAQ peptides experiments of different sample tags, Q-star mass spectrum identification and MASCOT library search. 153 proteins were identified as K-L 0 to K-L IV, 153 proteins were identified as K-L 0 to K-L II, and 145 proteins were identified as K-L II to K-L IV. iTRAQ technology may help to identify novel serum markers for early diagnosis of knee osteoarthritis, indicating iTRAQ technology for proteomics serum biomarkers of osteoarthritis study has good application prospect.%背景：在临床上对于早期膝关节骨性关节炎尚缺乏有效的诊断方式。蛋白质组学是从整体的角度分析细胞的所有蛋白质组成、表达水平及修饰状态的学科，可以了解蛋白与蛋白间的相互作用及联系，从而揭示蛋白功能与细胞生命活动的规律。 目的：通过对骨性关节炎K/L分级的各阶段的血清样本进行ITRAQ定量蛋白组学分析检测，以发现骨性关节炎各个时期的的潜在分子标志物。 方法：收集膝关节骨性关节炎患者60例，参照Kel gren-Lawrence(K-L)分级标准，分为K-L 0、Ⅱ、Ⅳ级的亚组，每个亚组随机选取10例男性与10例女性。去除血清高丰度蛋白，进行稳定同位素116、117、118的iTRAQ标记、反相色谱柱分离、质谱检测及Swissport数据库检索；再利用生物信息学软件进行分析。结果与结论：通过对不同样品的iTRAQ肽段实验标记、Q-star质谱鉴定和MASCOT搜库，共筛选出有意义差异蛋白169个，K-L 0级与K-L Ⅳ级差异蛋白153个；K-L 0级与K-L Ⅱ级差异蛋白153个；K-L Ⅱ级与K-L Ⅳ级差异蛋白145个。结果表明应用iTRAQ技术可筛选出与骨性关节炎发生发展的多个差异蛋白，提示iTRAQ技术对于骨性关节炎蛋白质组学血清生物标记物的研究有重要意义。
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