S-甲酰谷胱甘肽水解酶与类风湿性关节炎的相关性

发布时间：2018-02-01 18:32

本文关键词： 类风湿性关节炎 S-甲酰谷胱甘肽水解酶差异蛋白组学诊断标志　出处：《第三军医大学》2015年硕士论文　论文类型：学位论文

【摘要】：类风湿性关节炎(Rheumatoid arthritis,RA)是一种全身性自身免疫性疾病,是困扰大众健康的一大顽疾。在临床上,类风湿性关节炎有五大特点,致死、致残、痛苦、高花费、药物副作用大。随着疾病的发生发展,类风湿性关节炎患者如果没有得到早期诊断和及时的系统性治疗,还会引发除了关节损伤以外的各种并发症。并发症会发生在全身的各个重要脏器,严重时会对类风湿性关节炎患者的生命造成威胁。目前医学上,对其致病病理机制的了解尚未完全明确,临床上所运用的检测指标的特异性和敏感性不够高,致其治疗难度相应增加,所以发现和鉴定新的与疾病相关的特异性诊断标志物,是早期诊断类风湿性关节炎的必不可少的前提,并可能为类风湿性关节炎临床诊断和治疗提供新的靶点。为了找到新的与RA相关的自身抗原,我们首先采用二维凝胶电泳和免疫印迹法方法得到类风湿性关节炎患者血清蛋白图,和正常人血清比较寻找差异蛋白点。质谱测序和免疫印迹法证明S-甲酰谷胱甘肽水解酶(S-formylglutathione hydrolase,Es D)在患者血清中高表达、在正常人血清中则低表达。在此基础上,分别对早期类风湿性关节炎患者、晚期类风湿性关节炎患者、非类风湿性关节炎患者(其它自身免疫性疾病)和正常人血清通过ELISA的方法检测EsD水平,分析血清Es D水平与类风湿性关节炎的相关性,结果显示Es D在早期RA和晚期RA患者血清中表达水平明显高于非RA患者和正常人,表明EsD可能是RA潜在的一个诊断指标。进一步比较与类风湿性关节炎现有诊断指标的相关性,发现Es D血清水平与ESR、CCP、DAS28评分具有显著相关性(P0.05)而与RF无显著相关(P0.05),说明Es D血清表达水平与类风湿性关节炎疾病活动度呈正相关。最后我们选择8例治疗前与治疗后的类风湿性关节炎患者,发现Es D水平能够反映RA的治疗效果,该结果提示Es D亦可能同时作为类风湿性关节炎治疗效果的评价指标。研究结果显示:我们应用二维凝胶电泳和MALDI-TOF-MS质谱测序技术从类风湿性关节炎病人的血清中筛选出的差异蛋白点为EsD,在类风湿性关节炎病人的血清中检测到高水平表达。重要的是EsD水平与类风湿性关节炎疾病的临床指标如血沉(Erythrocyte sedimentation rate,ESR)、抗环瓜氨酸肽抗体、以及28关节评分(Disease activity score in 28 joints,DAS28)有明确的相关性。我们的结果提示Es D血清水平对类风湿性关节炎可能具有早期诊断价值,反映了疾病活动度;经过治疗后的RA患者血清中的Es D水平有明显下降,说明EsD能够反映RA的治疗效果。然而,Es D对早期RA的诊断价值需要更大的样本量验证,其在类风湿性关节炎疾病发生及进展中的诊断价值有待于进一步研究。
[Abstract]:Rheumatoid arthritis (RA) is a systemic autoimmune disease. Rheumatoid arthritis has five characteristics, death, disability, pain, high cost, drug side effects. With the development of the disease. If patients with rheumatoid arthritis do not get early diagnosis and timely systematic treatment, they will also cause a variety of complications other than joint injury. The complications will occur in all important organs of the whole body. At present, the pathogenetic mechanism of rheumatoid arthritis is not fully understood, and the specificity and sensitivity of clinical indicators are not high enough. As a result of the corresponding increase in the difficulty of treatment, so the detection and identification of new disease-related specific diagnostic markers, is an essential prerequisite for the early diagnosis of rheumatoid arthritis. And may provide a new target for the clinical diagnosis and treatment of rheumatoid arthritis. In order to find new antigens associated with RA. We first obtained the serum protein map of patients with rheumatoid arthritis by two-dimensional gel electrophoresis and Western blotting. The differential protein sites were identified by mass spectrometry sequencing and Western blotting. The results showed that S-formylglutathione hydrolase was detected by S-formylglutathione hydrolase. The high expression of es D was found in the serum of the patients and the low expression in the serum of the normal subjects. On this basis, the patients with early rheumatoid arthritis and the patients with advanced rheumatoid arthritis were treated respectively. The serum levels of EsD were measured by ELISA in patients with non-rheumatoid arthritis (other autoimmune diseases) and normal subjects, and the correlation between EsD levels and rheumatoid arthritis was analyzed. The results showed that the expression of es D in serum of early and late RA patients was significantly higher than that of non-RA patients and normal controls. It is suggested that EsD may be a potential diagnostic marker for RA. Further comparison with the existing diagnostic indexes of rheumatoid arthritis shows that the serum level of EsD is correlated with ESRCCP. DAS28 score was significantly correlated (P 0.05), but not to RF (P 0.05). The results showed that the expression of es D in serum was positively correlated with the activity of rheumatoid arthritis. Finally, we selected 8 patients with rheumatoid arthritis before and after treatment. It was found that the level of es D could reflect the therapeutic effect of RA. These results suggest that es D may also be used as an index to evaluate the therapeutic effect of rheumatoid arthritis. Two dimensional gel electrophoresis and MALDI-TOF-MS mass spectrometry sequencing techniques were used to screen the differential protein sites of EsD from the sera of patients with rheumatoid arthritis. High levels of expression were detected in the serum of patients with rheumatoid arthritis. It is important that the level of EsD and clinical indicators of rheumatoid arthritis such as erythrocyte sedimentation rate (ESR). Erythrocyte sedimentation rate. ESR, anti-cyclic citrullin-peptide antibody, and 28 joint score activity score in 28 joints. Our results suggest that the serum level of es D may have early diagnostic value for rheumatoid arthritis and reflect disease activity. After treatment, the serum EsD level of RA patients decreased significantly, indicating that EsD can reflect the therapeutic effect of RA. The diagnostic value of es D for early RA needs more sample size verification, and its diagnostic value in the pathogenesis and progression of rheumatoid arthritis needs to be further studied.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.22

【相似文献】