谷氨酰半胱氨酸连接酶在类风湿性关节炎中基因表达变化的研究

发布时间：2019-04-01 21:52

【摘要】：目的:类风湿性关节炎是一种常见的慢性自身免疫性关节疾病,其致病原因及病理过程一直是研究热点。为研究氧化应激状态特别是谷氨酰半胱氨酸连接酶对类风湿性关节炎疾病过程的影响,我们采用适当方法对谷氨酰半胱氨酸连接酶催化亚单位和调节亚单位在类风湿性关节炎患者体内的改变进行分析,以说明氧化应激在类风湿性关节炎发病中的意义。方法:采集类风湿性关节炎患者及健康对照清晨空腹静脉血,采用免疫比浊法检测RF、CRP、C3、C4、Ig G、Ig A、Ig M,仪器法检测血沉,ELISA法检测抗CCP抗体;使用Ficoll Hypaque淋巴细胞分层液密度梯度离心分离外周血单个核细胞,提取外周血淋巴细胞RNA,反转录,采用实时荧光定量PCR对谷氨酰半胱氨酸连接酶的催化亚基和调节亚基的基因表达水平进行分析,以溶解曲线验证实时荧光定量PCR实验扩增特异性。结果:类风湿性关节炎患者组实验室数据表现,抗CCP抗体、RF、CRP、血沉升高,C4降低,Ig G升高。与对照组相比较,类风湿性关节炎病人外周血淋巴细胞中的谷氨酰半胱氨酸连接酶催化亚基基因和调节亚基基因表达明显下降(谷氨酰半胱氨酸连接酶催化亚基基因:RA组1±0.018,对照组3.62±0.429;p0.01。谷氨酰半胱氨酸连接酶调节亚基基因:RA组1±0.03,对照组2.97±0.467;p0.01)。结论:谷氨酰半胱氨酸连接酶的催化亚基和调节亚基在类风湿性关节炎病人的疾病过程中发生明显的改变,其基因表达水平明显下降,说明谷胱甘肽代谢及机体的氧化应激状态可能在类风湿性关节炎的疾病过程中扮演重要的角色,并可将其作为研究类风湿性关节炎治疗的一个方向。
[Abstract]:Aim: rheumatoid arthritis (RA) is a common chronic autoimmune joint disease. In order to study the effects of oxidative stress, especially glutamylcysteine ligase, on the disease process of rheumatoid arthritis, We analyzed the changes of glutamylcysteine ligase catalytic subunit and regulatory subunit in patients with rheumatoid arthritis by appropriate methods to illustrate the significance of oxidative stress in the pathogenesis of rheumatoid arthritis. Methods: fasting venous blood samples were collected from patients with rheumatoid arthritis and healthy controls in the early morning. RF,CRP,C3,C4,Ig G, IgA, IgM, erythrocyte sedimentation rate (ESR) and anti-CCP antibody were detected by immunoturbidimetric assay, instrument method and anti-CCP antibody respectively. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation of Ficoll Hypaque lymphocytes, and reverse transcription of RNA, was extracted from peripheral blood lymphocytes. The gene expression levels of catalytic subunits and regulatory subunits of glutamylcysteine ligase were analyzed by real-time fluorescence quantitative PCR, and the specificity of real-time fluorescence quantitative PCR amplification was verified by dissolution curve. Results: the laboratory data of patients with rheumatoid arthritis showed that anti-CCP antibody, RF,CRP, erythrocyte sedimentation rate (ESR) increased and C4 decreased, Ig G increased. Compared with the control group, The expression of glutamylcysteine ligase catalytic subunit gene and regulatory subunit gene in peripheral blood lymphocytes of rheumatoid arthritis patients was significantly decreased (glutamylcysteine ligase catalytic subunit gene: RA group 1 卤0.018, P < 0.01). The control group was 3.62 卤0.429; P0.01. Glutamylcysteine ligase regulatory subunit gene: 1 卤0.03 in RA group and 2.97 卤0.467 in control group (P < 0.01). Conclusion: the catalytic subunits and regulatory subunits of glutamylcysteine ligase have obvious changes in the disease process of rheumatoid arthritis patients, and the gene expression level of glutamylcysteine ligase is significantly decreased. It is suggested that glutathione metabolism and oxidative stress may play an important role in the pathogenesis of rheumatoid arthritis and may be used as a research direction in the treatment of rheumatoid arthritis.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.22

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