类风湿关节炎患者自身反应性B细胞异常表达相关分子的检测和分析  

Aberrant expression of autoreactive B cells associated molecules in rheumatoid arthritis patients

作　　者：钱柳[1] 黄秋愉[1] 胡朝英[1,2] 程伟志[1] 苗平[1] 缪怡 余奇文[1] 张继英[1] 陈雪华[3] 何东仪 李卓凌 许荣 张冬青[1] 

机构地区：[1]上海交通大学医学院上海市免疫学研究所,上海200025 [2]徐汇区中心医院检验科,上海200031 [3]上海交通大学医学院附属瑞金医院外科上海消化外科研究所,上海200025 [4]上海市长宁区光华中西医结合医院检验科,上海200052

出　　处：《诊断学理论与实践》2011年第1期55-61,共7页Journal of Diagnostics Concepts & Practice

基　　金：国家自然科学基金项目(30471593);上海市科委重点项目(07JC14033和10JC1408500);上海市科委重点学科(外科学)开放课题(S30204-K01);上海市科委自然基金项目(10ZR1426100);上海市教科委重点项目(J50207);上海市免疫学研究所资助项目(08-A04)

摘　　要：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 correlation between autoreactive B cells and the expression of associated autoantibody and signal molecules in rheumatoid arthritis（RA） patients.Methods Flow cytometry was used to determine autoreactive B cells in peripheral blood of 22 RA patients and 10 normal controls and the expression of peripheral blood mononuclear cell（PBMC） Th2 in RA patients and osteoarthritis（OA） patients（n=10）.Fluorescent quantitative PCR（FQ-PCR） was used to detect the relative expression of extracellular signal regulated kinase（ERK） and focal adhesion kinase（FAK） genes in PBMC in RA patients and normal controls,and Western blot was used to analyze the phosphorylation pattern of NF-κB signaling pathway.By ELISA,serum and synovial fluid anti-collagen Ⅱ antibody level in RA and OA patients were determined.Meanwhile,the expressions of matrix metalloproteinase-3（MMP-3） and tissue inhibitor of matrix metalloproteinase-1（TIMP-1） in RA patients were determined.Results Compared with normal controls,autoreactive B cells with high expression of CD19＋ were present in the peripheral blood of RA patients.High relative expression of ERK gene and active phosphorylation of NF-κB signaling pathway were the causes of B cells aberrant function in RA patients.Compared with OA patients,anti-collagen Ⅱ antibody of RA patients in synovial fluid was increased.The expression of inflammatory factors MMP-3 and TIMP-1 of RA patients in synovial fluid was aberrant.Conclusions Autoreactive B cells and the aberrant expression of associated molecules might be another important factor mediating the pathogenesis of rheumatoid arthritis.

关 键 词：类风湿关节炎 自身反应性B细胞 细胞外信号调节激酶 抗Ⅱ型胶原抗体 基质金属蛋白酶-3 基质金属蛋白酶组织抑制因子-1 

分 类 号：R593.22[医药卫生—内科学]