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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。
Antibody against citrullinated protein is considered as the culprit of rheumatoid arthritis (RA), and the purpose of this study was to explore the pathogenesis of citrullinated fibronectin (cFn) in RA. The distribution of cFn in synovial tissue of RA patients was analyzed by immunohistochemistry and double immunofluorescence technique. Fibroblast-like synoviocytes (FLS) were isolated from synovial tissue of RA patients by fibronectin (Fn) and cFn treatment, and then apoptosis was detected by flow cytometry and TUNEL assay Happening. Real-time fluorescence quantitative PCR was used to detect the mRNA expression of survivin, Caspase 3 and cyclin-B1. The secretion of cytokines was measured by ELISA. Fn forms an extracellular citrullinated polymer in synovial tissue of RA patients. Fn induced apoptosis of RA-FLS, cFn inhibited the apoptosis of RA-FLS. FN in RA patients significantly increased the expression of caspase-3 and inhibited the expression of survivin and cyclin-B1. CFn significantly increased the expression of survivin and promoted the secretion of TNF-α and IL-1. During the development and progression of RA, cFn may exert its pathogenicity by inhibiting apoptosis and increasing the secretion of inflammatory cytokines.