Research of the pathogenesis of arthritis rheumatoid by micro RNA and msearch of the possibility of a novel diagnosis method for arthritis rheumatoid by micro RNA

类风湿关节炎(RA)是一种病因不明的慢性疾病，表现为一系列特征性的特征，包括白细胞向滑膜细胞的渗透和滑膜细胞的增生，导致血管疙瘩形成和关节破坏。这些细胞局部产生的细胞因子和趋化因子是类风湿关节炎许多病理和临床表现的原因。目前尚无早期诊断RA或预测其疗效的分子标记物。本研究的目的是通过分析一种新的RNA干扰因子miRNA(MiRNA)的表达，揭示RA的发病机制，探索一种新的诊断方法和治疗方法。我们发现5个miRNAs在RA-FLS中的表达强于骨关节炎FIS(0A-FLS)，而miR-124A是唯一一个在RA-FLS中显著低于OA-FLS的miRNA。将miR-124A前体基因导入RA-FLS，可显著抑制RA-FLS的增殖，并使细胞周期停滞于G1期，而对RA-FLS的凋亡率无明显增加。我们在CDK2和MCP1的3UTR区发现了一个假定的miR-124A结合位点，并且将miR-124A诱导到RA-FLS中显著抑制了CDK2和MCP1蛋白的产生。这些结果表明，RA-FLS中miR-124A的表达减少与RA的发病机制密切相关。另一个研究重点是鉴定外周血单个核细胞(PBMC)中的RA特异性miRNAs。我们一直在寻找RA特异的miRNA，初步数据显示，预期的miRNAs与RA-FLS完全不同，所有的研究费用都花在了购买化学试剂和一次性设备上。

Rheumatoid arthritis (RA) is a chronic disease of unknown etiology that presents a characteristic constellation of features that includes infiltrating leukocytes to synoviocytes and synoviocyte hyperplasia, resulting in pannus formation and joint destruction. The local production of cytokines and chemokines by these cells accounts for many of the pathological and clinical manifestations of RA. The molecular marker which could diagnose early RA or predict the response to currently used treatments does not exist.Our aim was to reveal the pathogenesis of RA by analyzing the expression of micro RNA (miRNA), a novel RNA interfence factor, and to exploit the possibility of inventing a novel diagnostic method and a novel therapy.One of the research foci was the identification of the specific miRNAs to RA fibroblast-like synoviocyte (RA-FIS). We found that five miRNAs were more strongly expressed in RA-FLS than in osteoarthritis FIS (0A-FLS), and miR-124a was the only significantly decreased miRNA in RA-FLS as compared to OA-FLS. The transfection of a precursor of miR-124a into RA-FLS suppressed their proliferation significantly and forced to stop the cell cycle at G1 phase with no significant increase of apoptosis to RA-FLS. We identified a putative consensus site for miR-124a binding in the 3UTR regions of CDK2 and MCP1 mRNA, and induction of miR-124a into RA-FLS significantly suppressed the production of CDK2 and MCP1 proteins. These results suggest that decreasing miR-124a expression in RA-FLS is deeply involved in the pathogenesis of RA.Another research focus was to identify the RA-specific miRNAs in peripheral blood mononuclear cells (PBMC). We have been searching for RA-specific miRNA in PBMC, and preliminary data show that totally different sets of miRNAs from that of RA-FLS are anticipated.All research expenses were spent to buy chemical reagents and disposable equipments.