CTGF蛋白促进类风湿性关节炎血管翳生成的转录调控机制研究

类风湿性关节炎（RA）是一种病因复杂的自身免疫性疾病。已发现滑膜成纤维细胞的过度增殖及血管翳的形成对RA疾病进程至关重要，但血管翳生成的机制仍不清楚。申请人前期发表在Arthritis & Rheumatism上的蛋白质组学及差异蛋白功能研究结果发现CTGF蛋白在RA滑膜成纤维原代培养细胞高表达并与血管生成相关。本项目将通过体内和体外实验进一步验证CTGF蛋白对RA血管翳生成的促进作用；同时应用生物信息学预测与CTGF启动子结合的转录因子，通过双荧光素酶报告基因，EMSA和染色体免疫共沉淀等技术研究CTGF蛋白的转录调控机制，并通过体内实验进一步验证其对RA血管翳生成的影响；通过ELISA实验大样本验证CTGF蛋白在RA病人及正常人群中血清，滑液及滑膜病例标本中的差异表达。旨在明确CTGF蛋白促进RA血管翳生成的转录调控机制，为开发新的生物标志物用于RA的临床诊断及药物干预靶点提供理论依据

Rheumatoid arthritis (RA) is an autoimmune disease for complex etiology, It has been found that the excessive proliferation of synovial fibroblast and formation of pannus is critical to RA disease processes. but the mechanism of pannus generation remains unclear. Our previous studies of proteomics and the functional of differentially expressed proteins found that CTGF protein is over expressed in RA FLS cells which correlated with blood vessels formation. The further studies need to be done, include the mechanisms of CTGF regulating RA pannus generation by experiments in vivo and in vitro; while bioinformatics predict the transcription factors binding with CTGF promoter, and the mechanism study of transcriptional regulatory of CTGF protein by the dual -luciferase reporter system, EMSA and chromosome immunoprecipitation, and further validate the effects generated by the RA pannus in vivo; idetify the differentially expression of CTGF protein in serum, synovial fluid and synovium of RA and normal samples by ELISA. The aim of this study is to clarify the mechanism of pannus formation for diagnosis of RA and new drugs targets discovery.