瓜氨酸化纤维连接蛋白对类风湿性关节炎成纤维样滑膜细胞的凋亡及促炎症因子分泌的影响  

Effects of citrullinated fibronectin on the apoptosis of fibroblast-like synoviocytes and the secretion of pro-inflammatory cytokines in rheumatoid arthritis

作　　者：郭金虎[1] 宗明[1] 张慧[1] 虞姗姗 龚如涵 范列英[1] 

机构地区：[1]同济大学附属东方医院检验科,上海200120

出　　处：《现代免疫学》2014年第2期103-108,共6页Current Immunology

基　　金：国家自然科学基金面上项目(81373203);上海市科委生物医药领域重点项目(11DZ1973802);上海市优秀学术带头人计划(12XD1404300)

摘　　要：针对瓜氨酸化蛋白的抗体被认为是诱发类风湿性关节炎(rheumatoid arthritis,RA)的元凶,本研究旨在探讨瓜氨酸化纤维连接蛋白(citrullinated fibronectin,cFn)在RA中的致病机理。通过免疫组化和双重免疫荧光技术分析cFn在RA患者的滑膜组织中的分布情况。用纤维连接蛋白(fibronectin,Fn)以及cFn处理从RA患者的滑膜组织中分离得到成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)后,再通过流式技术和TUNEL法检测细胞凋亡情况。实时荧光定量PCR检测survivin,Caspase 3,cyclin-B1的mRNA表达情况。用ELISA法测定细胞因子的分泌。在RA患者的滑膜组织中Fn形成胞外瓜氨酸化聚合物。Fn诱导了RA-FLS的凋亡,cFn抑制了RA-FLS的凋亡。在RA患者的FLS中,Fn显着增加了caspase-3的表达,抑制了survivin和cyclin-B1的表达;cFn显著增加了survivin的表达,促进了TNF-α和IL-1的分泌。在RA的发生和发展过程中,cFn可能通过抑制细胞凋亡并且增加炎症因子的分泌来发挥其致病性。This study aims to investigate the pathogenesis of eitrullinated fibronectin（cFn） in rheumatoid arthritis （RA）. Through immunohistoehemical and double immunofluorescence analysis we get the distribution of fibronectin （FN） and eFN in RA patients＇ synovial tissue. Fibroblast-like synoviocytes （FLS） were isolated from RA patients and treated with Fn or cFn. Flow cytometry and TUNEI. assay were used to detect apoptosis. The expression of survivin, caspase-3, and cyclin-B1 was de- tected by real-time PCR. The secretion of proinflammatory cytokines was measured by ELISA. eFn formed extraeeltular aggre- gates in synovial tissues of RA patients. Fn induced apoptosis of RA FLS while cFn inhibited the apoptosis of RA FLS. Fn sig- nificantly increased the expression of caspase-3 and decreased the expression of survivin and cyclin-B1 in FLS from RA patients. cFn significantly increased the expression of survivin in RA FLS. Furthermore, cFn increased the secretion of TNF-α and IL-1 by FIRS. The results suggest that cFn plays a potential pathophysiologic role in RA by inhibiting apoptosis and increasing proin- flammatory cytokine secretion by FLS.

关 键 词：类风湿性关节炎 成纤维样滑膜细胞 瓜氨酸纤维连接蛋白 纤维连接蛋白 

分 类 号：R392.12[医药卫生—免疫学]