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目的:分析类风湿关节炎(rheumatoid arthritis,RA)患者自身反应性B细胞及与之功能相关的自身抗体和信号分子的表达。方法:采用流式细胞仪分析22例RA患者及10例正常对照者的外周血自身反应性B细胞比率及RA和骨关节炎(OA)患者(n=10)外周血单个核细胞(PBMC)Th2的表达。用荧光定量PCR(FQ-PCR)检测RA患者及正常对照者PBMC细胞外信号调节激酶(ERK)、黏着斑激酶(FAK)基因的相对表达量,并采用蛋白印迹法检测NF-κB信号通路的磷酸化格局;用ELISA法检测和分析RA及OA患者血清及关节滑膜液抗Ⅱ型胶原抗体水平,同时检测RA患者血清及关节滑膜液中基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶组织抑制因子-1(TIMP-1)的表达状况。结果:相对于正常对照者,RA患者外周血中存在高表达CD19+的自身反应性B细胞;同时其PBMC中ERK基因相对表达量提高、NF-κB信号通路磷酸化活跃。相对于OA患者,RA患者关节滑膜液中抗Ⅱ型胶原抗体水平显著升高。RA患者关节滑膜液中MMP-3及TIMP-1等水平较其自身血清中升高。结论:RA患者自身反应性B细胞及相关分子的异常表达可能是介导RA发病的又一重要因素。
OBJECTIVE: To analyze the expression of autoantibodies and signal molecules related to auto-reactive B cells and their functions in patients with rheumatoid arthritis (RA). Methods: Peripheral blood autoreactive B cell ratio and peripheral blood mononuclear cells (PBMCs) in RA and osteoarthritis (OA) patients (n = 10) were analyzed by flow cytometry in 22 RA patients and 10 normal controls. Th2 expression. The relative expression of extracellular signal-regulated kinase (ERK) and focal adhesion kinase (FAK) in peripheral blood mononuclear cells of RA patients and normal controls were detected by real-time quantitative polymerase chain reaction (FQ-PCR) The levels of MMP-3 and MMP-3 in serum and synovial fluid of RA patients were detected by ELISA. Metalloproteinase tissue inhibitor of metalloproteinase-1 (TIMP-1) expression status. Results: Peripheral blood mononuclear cells with auto-reactive B cells were highly expressed in RA patients compared with normal controls. The relative expression of ERK gene in PBMCs was increased and phosphorylation of NF-κB signaling pathway was active. Relative to OA patients, RA patients with synovial fluid anti-type Ⅱ collagen antibody levels were significantly higher. RA patients with synovial fluid MMP-3 and TIMP-1 levels higher than its own serum. Conclusion: Abnormal expression of autoreactive B cells and related molecules may be another important factor in the pathogenesis of RA.