An Yuan, Pu Hai, He Si-zhi, Li Zhan-guo
Department of Rheumatology and Immunology, People's Hospital, Peking University, Beijing 100044, China.
Zhonghua Yi Xue Za Zhi. 2005 May 18;85(18):1261-5.
To discover the biomarkers of early rheumatoid arthritis (RA) patients and healthy person were analyzed by proteomic methods to discover serum biomarkers.
Samples of peripheral blood were collected form 10 newly diagnosed active RA patients, 5 males and 5 females, aged 54.3 +/- 12.78, with a disease course of 4.08 +/- 1.9 months, and 10 age and sex matched healthy persons. The samples were divided into 10 groups containing 1 sample of patient serum and 1 sample of healthy serum to undergo. High-molecular-weight protein (HMWP) was enriched by depletion of albumin with HiTrap Blue column and depletion of immunoglobulin G with HiTrap rProtein A column. Low-molecular-weight protein (LMWP) was enriched by C18 absorbent binding and elution. After comparative proteomic analysis, the different protein spots were identified using matrix-assisted laser desorption ionization-time of flight-ionization-mass spectrometry. (MALDI-TOF-MS).
Marrow-related protein (MRP) 14, a protein of the S100 family was 100% positive in the RA patient sera and 100% negative in the sera of the healthy controls. MRP8, another protein of the S100 family, was 100% positive in the RA patient sera and 50% positive in the sera of the healthy controls. Ubiquitin was 90% positive in the RA patient sera and 10% positive in the sera of the healthy controls. The levels of apolipoprotein A-I (ApoA-I), serum amyloid A1 and A2 (SAA1/SAA2), and transthyretin of the RA patients were all significantly higher than those of the healthy controls.
MRP14/MRP8, SAA1/SAA2 and ubiquitin may play important roles in development of RA and their determination may benefit early diagnosis, evaluation of disease activity and investigation of new therapy targets.
通过蛋白质组学方法分析早期类风湿关节炎(RA)患者和健康人的样本,以发现血清生物标志物。
收集10例新诊断的活动期RA患者的外周血样本,其中男性5例,女性5例,年龄54.3±12.78岁,病程4.08±1.9个月,以及10例年龄和性别匹配的健康人样本。将样本分为10组,每组包含1例患者血清样本和1例健康血清样本。通过HiTrap Blue柱去除白蛋白和HiTrap rProtein A柱去除免疫球蛋白G来富集高分子量蛋白(HMWP)。通过C18吸附剂结合和洗脱来富集低分子量蛋白(LMWP)。经过比较蛋白质组学分析后,使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定不同的蛋白斑点。
S100家族的一种蛋白骨髓相关蛋白(MRP)14在RA患者血清中100%呈阳性,在健康对照血清中100%呈阴性。S100家族的另一种蛋白MRP8在RA患者血清中100%呈阳性,在健康对照血清中50%呈阳性。泛素在RA患者血清中90%呈阳性,在健康对照血清中10%呈阳性。RA患者的载脂蛋白A-I(ApoA-I)、血清淀粉样蛋白A1和A2(SAA1/SAA2)以及转甲状腺素蛋白水平均显著高于健康对照。
MRP14/MRP8、SAA1/SAA2和泛素可能在RA的发生发展中起重要作用,对它们的检测可能有助于早期诊断、疾病活动度评估及新治疗靶点的研究。