【摘要】 目的:采用类风湿性关节炎(RA)患者滑膜细胞的体外培养,研究基质蛋白Cyr61在RA滑膜细胞增殖中的作用及其机制。方法:通过Real-time PCR、Western blot和免疫组化检测RA病人的滑膜组织和细胞中Cyr61的表达情况;用3H-TdR掺入法检测滑膜液(SF)对滑膜细胞增殖的影响;用ELISA方法检测RA患者滑膜液中Cyr61蛋白的水平。结果:RA病人的滑膜组织和细胞中高表达Cyr61;SF能刺激滑膜细胞发生明显增殖;且RA患者滑膜液中含高浓度的Cyr61蛋白。用SiRNA干扰技术抑制滑膜细胞中Cyr61基因表达,再加入滑膜液后,则滑膜细胞增殖明显降低。同时,将SF与anti-Cyr61抗体共同孵育后再刺激滑膜细胞,FLS也不再发生明显增殖。进一步研究滑膜液中与上调Cyr61表达有关的炎症细胞因子,发现SF中IFNγ-和TNFα-具有上调Cyr61蛋白表达的作用。结论:Cyr61蛋白是促进滑膜细胞增殖的重要调控基因;RA患者滑膜液中含有高浓度的炎症因子IFNγ-和TNFα-,通过上调Cyr61蛋白表达而促进滑膜细胞增殖,可能是促进RA病理性滑膜增生的重要因素之一。更多还原

【Abstract】 Objective:To investigate the effect of Cyr61 on the proliferation of Fibroblast-like Synoviocytes(FLS) in rheumatoid arthritis(RA).Methods:Cyr61 expression in synovial tissues(ST) and FLS was examined using Real-time PCR,Western blot and immunohistochemistry simultaneously.FLS were isolated from synovial tissue of RA patients and cultured in vitro.The proliferation of FLS stimulated with synovial fluid(SF) was determined by 3 H-TdR incorporation.Cyr61 protein level in RA SF was detected by ELISA.Results:Cyr61 was over expressed in ST,FLS of RA patients while hardly examined in normal individuals and osterarthritis(OA) patients.Meanwhile,Cyr61 protein level was elevated in SF,which promoted the proliferation of RA FLS.This proliferation was abrogated by knockdown the Cyr61 gene of FLS or neutralizing monoclonal antibody against human Cyr61.Moreover,inflammatory factor IFN-γ and TNF-α up-regulated the expression of Cyr61.Conclusion:These results indicate that the elevated level of IFN-γ and TNF-α in RA SF can promote the proliferation of the FLS derived from RA patients by increasing expression of Cyr61,suggesting that Cyr61 may play an important role in the development of RA.更多还原