Henrotin Yves, Gharbi Myriam, Mazzucchelli Gabriel, Dubuc Jean-Emile, De Pauw Edwin, Deberg Michelle
Bone and Cartilage Research Unit, University of Liège, Liège, Belgium.
Arthritis Rheum. 2012 Jul;64(7):2260-7. doi: 10.1002/art.34392.
This study was undertaken to identify new biomarkers of osteoarthritis (OA) by proteomics analysis and to develop specific immunoassays to detect and quantify them.
Proteomics analysis was performed in urine samples from 10 women (mean±SD age 76.0±5.0 years) undergoing knee replacement surgery due to severe OA and 5 healthy women (mean±SD age 25.6±2.6 years). Protein content was analyzed by 2-dimensional differential gel electrophoresis. Protein spots that exhibited an OA:control abundance ratio of ≥1.5 were identified by mass spectrometry. Specific enzyme-linked immunosorbent assays were developed and validated in serum obtained from 236 healthy subjects ages 20-64 years and from 76 patients with severe radiologic knee OA (mean±SD age 68.8±11.9 years). Immunohistochemical analysis was performed on articular cartilage from tibial plateaus.
Thirteen proteins within spots that were significantly modified between groups were identified. Two peptides of fibulin 3, named Fib3-1 and Fib3-2, were of particular interest. Two antisera directed against these peptides were used to develop immunoassays. Compared with age-matched healthy subjects, median levels of serum Fib3-1 and Fib3-2 were elevated in OA patients (54.6 pM versus 85.1 pM [P<0.0001] and 144.4 pM versus 191.4 pM [P<0.0001], respectively). Using area under the receiver operating characteristic curve analysis, we demonstrated that Fib3-1 and Fib3-2 levels discriminate between OA and normal populations. Immunostaining revealed the presence of Fib3-1 and Fib3-2 in chondrocytes and in the extracellular matrix of the superficial layer of the fibrillated cartilage.
Our findings indicate that Fib3-1 and Fib3-2 are potential biochemical markers for the diagnosis of OA.
本研究旨在通过蛋白质组学分析鉴定骨关节炎（OA）的新生物标志物，并开发特异性免疫测定方法以检测和定量这些标志物。
对10名因严重OA接受膝关节置换手术的女性（平均±标准差年龄76.0±5.0岁）和5名健康女性（平均±标准差年龄25.6±2.6岁）的尿液样本进行蛋白质组学分析。通过二维差异凝胶电泳分析蛋白质含量。质谱鉴定OA与对照丰度比≥1.5的蛋白质斑点。在236名年龄20 - 64岁的健康受试者和76名严重膝关节OA放射学患者（平均±标准差年龄68.8±11.9岁）的血清中开发并验证了特异性酶联免疫吸附测定。对胫骨平台的关节软骨进行免疫组织化学分析。
鉴定出组间有显著修饰的斑点内的13种蛋白质。纤维连接蛋白3的两种肽段，命名为Fib3 - 1和Fib3 - 2，特别受关注。针对这些肽段的两种抗血清用于开发免疫测定。与年龄匹配的健康受试者相比，OA患者血清Fib3 - 1和Fib3 - 2的中位数水平升高（分别为54.6 pM对85.1 pM [P<0.0001]和144.4 pM对191.4 pM [P<0.0001]）。使用受试者工作特征曲线下面积分析，我们证明Fib3 - 1和Fib3 - 2水平可区分OA和正常人群。免疫染色显示Fib3 - 1和Fib3 - 2存在于软骨细胞和原纤维软骨表层的细胞外基质中。
我们的研究结果表明，Fib3 - 1和Fib3 - 2是诊断OA的潜在生化标志物。
