摘要
目的探讨类风湿关节炎(RA)早期患者血清蛋白质与正常人的差异。方法(1)选取10例病程1年内的早期RA患者血清以及10名正常对照。(2)血清样品处理高分子量蛋白质通过亲和层析以HiTrapTMBlue柱亲和除去白蛋白随后以HiTrapTMrPriteinA柱亲和除去IgG进行富集。低分子量蛋白质通过C18吸附剂吸附梯度洗脱处理血清进行富集。(3)通过双向电泳进行差异比较,利用基质辅助激光解吸电离飞行时间质谱(MALDI TOF MS)鉴定差异蛋白点。结果通过比较RA患者与正常对照血清的高分子量及低分子量蛋白质双向电泳,发现了RA患者中存在多个表达异常蛋白质点,差异蛋白质点质谱鉴定结果(1)RA患者血清中髓系相关蛋白(MRP)14阳性比例为10/10,正常对照均阴性;RA中MRP8阳性比例为10/10,正常对照为5/10;RA中泛素阳性比例为9/10,正常对照为1/10。(2)RA患者中载脂蛋白A I(ApoA I)、血清淀粉样蛋白(SA)A1、A2和转甲状腺蛋白含量明显高于正常人。结论MRP14/MRP8,SAA1/SAA2和泛素可能在RA致病过程中发挥了作用。
Objective To discover the biomarkers of early rheumatoid arthritis (RA) patients and healthy person were analyzed by proteomic methods to discover serum biomarkers.Methods 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 ~TM Blue column and depletion of immunoglobulin G with HiTrap^TM 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).Results 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.Conclusion 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.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2005年第18期1261-1265,共5页
National Medical Journal of China
