新型自身抗原RBP1类似蛋白Rbik与汉防己碱治疗高原缺氧地区类风湿性关节炎相关性研究

Rheumatoid arthritis (RA) is an autoimmune disease characterized by long-term inflammation and resultant destruction of multiple joints. The incidence of RA distributed at Tibet Plateau is comparatively higher than that of inland, which is determined by their nature conditions. The Synovial oxigen level in RA patients is relatively lower compare to that in normal people. Thus, we conclude that hypoxia is derectly related to RA progression. In joint spaces of patients with rheumatoid arthritis (RA), persistently presented antigens appear to play a major role in sustained inflammation by continuously stimulating T and B cells. Synovium seems to supply most of such antigens. We cloned RBP1-like protein from synovial tissue, named Rbik, as an autoantigen in RA. Rbik has 36.5% homology with retinoblastoma binding protein 1 (RBP1) which associates with retinoblastoma (RB) tumor suppressor protein and is thought to thereby modulate the gene repression activity of RB. Serum IgG class antibodies will be assayed by immunoblotting with polyhistidine-tagged full-length Rbik protein, which will be expressed by E. coli and purified by a cobalt-based metal affinity chromatography method. The function of Rbik remains to be clarified. we report that association of RA with drugs: Correlational analysis of RA between new autoantibodies to RBP1-like Protein (Rbik) and Sinaomenine.

类风湿性关节炎是一种病因尚未明了的全身性自身免疫性疾病。西藏高原的自然条件决定了高原地区类风湿性关节炎发病率远较内地高。RA患者滑膜液中氧含量水平相对正常人有所降低。缺氧与RA病理进程直接关联。本课题组运用克隆技术得到三个新型cDNA,其中的两个抑制类风湿性关节炎，且两者的抗体出现都与疾病活动度有直接联系。第三个91kDa新型蛋白，与RBP1有36.5%的氨基酸序列相同，因此命名RBP1 like Protein(Rbik)。本课题拟利用基因工程构建重组Rbik表达载体，精致蛋白，以此为抗原，western blot和ELISA法检测，并导入滑膜细胞系，研究滑膜新型抗原Rbik在类风湿性关节炎发病机制中的可能作用以及在缺氧调节机制中的影响，同时探讨了常用的治疗类风湿性关节炎中药单体汉防己碱在治疗类风湿性关节炎中与新型抗原Rbik的相关性，开创一个新的中药治疗观测指标和机理解析。

类风湿关节炎（RA）是一种自身免疫性疾病，其特征是长期炎症和多关节破坏。但其发病原因尚不清楚。本课题组运用克隆技术发现新型自身抗原，视黄醇结合蛋白1类似蛋白（RBP1 like protein，Rbik）。在大肠杆菌中表达Rbik，以此作为抗原，运用蛋白质印迹法（Western blotting）法检测患者血清中的抗Rbik抗体。分析Rbik蛋白与临床指标的相关性，探讨抗Rbik抗体的临床运用。结果显示，类风湿性关节炎（RA）患者血清中抗Rbik抗体阳性率41%，系统性红斑狼疮（SLE）11%，干燥综合征（SS）20%，正常人全部为阴性。RA患者血清抗Rbik抗体阳性率显著高于SLE，SS和正常人。因此我们进一步分析了抗Rbik抗体与RA病程的相关性。C反应蛋白CRP），红细胞沉降率（ESR），类风湿性因子（RF），基质金属蛋白酶3(MMP3)这些RA活动度指标均未得到有意义的相关性结果。但是，根据RF值分为三组，RF<10组，100>RF≥10组，RF≥100组。CRP，ESR在RF<10时，抗Rbik抗体阳性组较低。MMP3在RF≥100时，抗Rbik抗体阳性组较高。当RF≥100时，IgG在抗Rbik抗体阳性组显著升高。目前临床诊断抗CCP抗体具有较高的敏感度和特异性，但是，抗Rbik抗体可能在RA的预后，关节损伤的预测，以及联合抗CCP抗体提高诊断敏感度和特异性中起阳性作用。我们将制作RA联检试剂盒，主要检测Rbik和其他6个自身抗体，将彻底解决检测敏感度低，单因子检测平台杂乱，检测标准不统一的问题。