摘要
目的类风湿关节炎(RA)是一种典型的Ⅲ型超敏反应,可在滑膜组织中发现大量的免疫复合物及补体的沉积,但具体的致病机制尚不清楚。文中旨在探究RA血清免疫复合物中抗原谱的表达情况。方法血清样本取自2013年1月至2015年12月南京军区南京总医院门诊和住院RA患者共55例为RA组。RA组分为2个亚组,其中抗CCP抗体(+)RA亚组41例,抗CCP抗体(-)RA亚组14例。同时选取SLE患者41例血清样本为疾病对照组,健康体检人群41例血清样本为健康对照组。用PEG沉淀法从4组混合血清中提取免疫复合物,然后通过免疫共沉淀、胶内酶解后进行质谱分析。用Vennny2.1.0进行各组之间总蛋白差异比对,结果以RA组蛋白FOT值大于疾病对照组、健康对照组FOT值的2倍即判定为上调蛋白,小于疾病对照组、健康对照组FOT值0.5倍判定为下调蛋白。利用STRING软件对差异蛋白进行进一步功能分析。结果在RA患者血清免疫复合物中共鉴定出277种蛋白,其中抗CCP抗体(+)RA亚组有162种蛋白,抗CCP抗体(-)RA亚组有248种蛋白。相对疾病对照组与健康对照组,仅在RA组出现共有129种蛋白,其中,抗CCP抗体(+)RA亚组有38种蛋白,抗CCP抗体(-)RA亚组有109种蛋白,2亚组共有18种蛋白。RA组与对照组共有蛋白中,抗CCP抗体(+)RA亚组有2种蛋白上调,抗CCP抗体(-)RA亚组有11种蛋白上调。下调蛋白中,抗CCP抗体(+)RA亚组有17种蛋白表达下调,抗CCP抗体(-)RA亚组共计21种蛋白表达下调。结论抗CCP抗体(-)RA亚组相对抗CCP抗体(+)RA亚组鉴定出更多的差异蛋白,差异蛋白的鉴定为探究RA发病机制寻找新的生物学标记物提供新的思路和方向。
Objective Rheumatoid arthritis（ RA） is a typical type Ⅲ hypersensitivity with a large number of immune complexes（ IC） and complement deposits in the synovial tissue,but its specific pathogenesis is not yet clear. This article was to explore the expression of the antigenic profile of serum ICs in RA. Methods ICs were isolated from the serum of 55 patients with RA（ 41 cases of anti-CCP antibody [＋] and 14 cases of anti-CCP antibody [-]）,41 with systemic lupus erythematosus（ SLE）,and another 41 healthy controls by polyethylene glycol（ PEG） precipitation,separated by immunoprecipitation,digested with trypsin in gel,and then subjected to mass spectrometry for identification. The levels of total proteins were compared among different groups using Vennny 2. 1. 0. The protein expression was considered to be up-regulated when the total protein level of the RA group was 〉2 times and down-regulated when it was 〈0.5 times that of the control. Further functional analysis was performed on the differential proteins in RA using the STRING software. Results Totally,277 proteins were identified in the serum ICs of the RA patients,including 162 in the anti-CCP（ ＋） and 248 in the anti-CCP（-） RA group. Compared with the SLE and healthy control groups,only 129 proteins were found in the RA patients,including 38 in the anti-CCP（ ＋）,109 in the anti-CCP（-）RA group,and 18 in both the two groups. Among the proteins identified in the RA patients and healthy controls,2 and 11 were up-regulated while 17 and 21 down-regulated in the anti-CCP（ ＋） and anti-CCP（-） RA group,respectively. Conclusion More differentially expressed proteins were identified in the anti-CCP（-） than in the anti-CCP（ ＋） RA patients. The identification of differentially expressed proteins provides a new idea and direction for the investigation of the pathogenesis and new biomarkers of RA.
出处
《医学研究生学报》
CAS
北大核心
2017年第5期495-501,共7页
Journal of Medical Postgraduates
基金
国家自然科学基金(81071419
81470071)
江苏省科技支撑计划-社会发展项目(BE2011786)
关键词
类风湿关节炎
质谱
免疫复合物
Rheumatoid arthritis
Mass spectrometry
Immune complex
