泛素连接酶Pellino1调控pDCs的I型干扰素通路在SLE自身免疫性炎症中的作用及机制研究

Plasmacytoid Dendritic cells(pDCs) which producing very large amounts of type I interferon, have a important role in immune disorder of systemic lupus erythematosus (SLE), but the mechanism remains unclear. Ubiquitin ligase Pellino1(Peli1) is the key molecular in many signaling pathways, such as TLR/IL-1R, TCR-CD28. And our previous study found that Peli1 gene polymorphisms are closely associated with SLE. Our hypothesis is that Peli1 is the key molecule in type I IFN signaling pathway of pDCs, which involved in SLE autoimmune inflammation. In this study, we will build Peli1 RNA interference plasmid, separation lupus mice spleen pDCs, and then detect the expressions of Peli1 and type I IFN-related cytokines such as IRAK1, IRF7 before and after Peli1 RNA interference, in order to demonstrate that Peli1 plays an important role in pDCs dysfunction of SLE. This study is expected to provide a new target for the treatment of SLE.

浆细胞样树突状细胞（pDCs）产生大量I型干扰素（IFN）造成免疫紊乱是系统性红斑狼疮（SLE）发生发展的重要环节，但机制尚不清楚。泛素连接酶Pellino1（Peli1）是免疫细胞中TLR/IL-1R、TCR-CD28等信号途径的关键分子，同时我们前期研究也发现Peli1编码基因多态性与SLE密切相关。我们推测：Peli1是pDCs I型IFN信号途径的关键分子，参与了SLE自身免疫性炎症的形成。本课题拟构建Peli1 RNA干扰质粒，体外分选狼疮鼠脾脏pDCs，干扰Peli1表达后用TLR配体刺激，分子生物学技术检测Peli1及相关信号分子IRAK1、IRF7、IFNα等表达；体内实验则行ANA滴度和肾脏病理检查，流式分析pDCs数量和表型。本研究有望阐明Peli1在pDCs中对I型IFN途径的调控作用及对SLE发病机制的影响，为SLE临床治疗提供新的思路和干预靶点。

由于自我更新、多重分化能力和降低免疫原性, 骨髓间充质干细胞(BMSC)被认为是一种有前途的替代细胞,用于治疗系统性红斑狼疮(SLE)。最近的证据表明,SLE BMSCs具有凋亡、细胞循环阻滞和衰老表型,可能导致SLE的进展。本研究使用CIBERSORT方法，从GSE50772数据集中估算SLE患者的pbmc和健康对照组中各种免疫细胞的丰度。用WGCNA筛选免疫细胞相关共表达模块，用LASSO算法确定相关特征基因。采用ELISA法测定5例SLE患者和5例对照组的血清炎症趋化因子。分离BMSCs，通过RT-qPCR和western blotting检测TK1在BMSCs中的表达。转染TK1过表达和TK 1沉默的SLE BMSCs，流式细胞术检测细胞凋亡和细胞周期。western blotting检测细胞凋亡、细胞周期和衰老相关蛋白。在SLE的BMSCs中,TK1表达被证明是上调的。TK1–过表达的SLE的BMSCs呈现增强IL-1β表达、凋亡、G1抑制和衰老表型,并在SLE的TK1-沉默 BMSCs中结果相反。因此,我们的发现可能会为SLE提供潜在的新的治疗靶点。