Cyr61促进类风湿关节炎发病的机制研究

In the pathogenesis of Rheumatoid Arthritis, both of macrophages and Cyr61 play important roles and they have tight connections with each other. Pyroptosis, one of the common death forms of macrophages, can induce inflammation. Cyr61 is involved in cell apoptosis, which can convert to pyroptosis. In the previous study, we found that the serum concentrations of Cyr61 and its receptor Syndecan4 in RA were higher than those in healthy controls, and both of them were related with disease activity of RA. We also identified pyroptosis in fibroblast of RA synovium. In the current study, we plan to 1) analyze the association of pyroptosis associated molecules and RA disease activity; test 2) the impact of ectogenic Cyr61 and knock-down of Cyr61 on RA macrophage pyroptosis related molecules; 3) the change of pyroptosis associated molecules after blocking associated receptors and clearing ROS; 4) the influence of Cyr61 or anti-Cyr61 monoclone antibody on ROS and pyroptosis associated molecules of mice BMDM; 5) the influence of Cyr61 on the mice arthritis severity of CIA mice after depletion of macrophage. We aim to verify the hypoptosis that Cyr61 can promote RA by inducing macrophage pyroptosis and elucidate possible receptors on macrophage in the process, in order to seek new therapy strategies of RA.

在类风湿关节炎（RA）的发病中，巨噬细胞及Cyr61均起重要的促进作用，二者关系密切。焦亡是巨噬细胞常见的死亡方式，可诱导炎症。Cyr61参与的细胞凋亡，凋亡可向细胞焦亡转化。我们发现RA血清中Cyr61及其受体Syndecan4高于正常人，与疾病活动度相关，RA患者滑膜中存在焦亡现象。本项目拟通过（1）分析RA患者巨噬细胞焦亡通路相关分子与疾病活动度的关系；检测（2）外源性加入或敲低巨噬细胞Cyr61对RA患者巨噬细胞焦亡通路相关分子的影响；（3）封闭可能的受体及清除活性氧簇（ROS）后RA巨噬细胞焦亡通路分子的变化；（4）Cyr61单抗及外源性Cyr61对关节炎小鼠BMDM中ROS及焦亡通路分子的影响；（5）清除巨噬细胞后，Cyr61对小鼠关节炎表型的影响，证实Cyr61通过诱导RA巨噬细胞的焦亡促进RA的发病，寻找此过程涉及的Cyr61受体，为寻求RA的新治疗靶点提供理论依据。

类风湿关节炎滑膜成纤维样细胞（RA-FLS）通过其促炎及骨破坏作用参与类风湿关节炎（RA）发病，但其机制尚不清楚。研究表明，高半胱氨酸蛋白61（cyr61）和内质网应激/自噬通路二者均与RASFs异常增殖和促炎作用相关。我们前期工作发现，cyr61能够激活RASFs内质网应激/自噬通路，尤其是内质网感受器蛋白IRE1/p-JNK/自噬通路。本项目在滑膜组织、体外细胞水平，通过RA-FLS原代细胞培养、RT-PCT、Western-blot、慢病毒siRNA-Cyr61转染等方法，明确在 RA 患者的滑膜成纤维细胞中存在细胞焦亡；但Cyr61并不促进RA 患者滑膜成纤维细胞的焦亡；明确在RA 患者的滑膜成纤维细胞中存在自噬现象，Cyr61通过ROS/Hif-1α/BNIPs通路促进RA患者滑膜成纤维细胞的自噬，并进一步促进RA-FLS的迁移及侵袭功能。Cyr61是RA发病过程中的重要细胞因子，可能作为将来RA治疗的靶点。