探索儿童系统性红斑狼疮MicroRNA-525-5p高表达导致病情活动的作用机制

Some miRNAs can cause the disorder of cytokine expression and lead to disease activity of pSLE, as cytokines disorder is one of the important mechanisms. Our previous study found that the expressionlevel of miR-525-5p and disease activity of pSLE are positive correlation. On the basis of the research above, this project firstly intends to use overexpression or silencing of miR-525-5p molecular in the peripheral blood mononuclear cells, activated T cells, 293A cells and the lupus murine model to prove the molecular mechanism how miR-525-5p causes the disorder of RANTES and other cytokines.Secondly, we verified the target gene of miR-525-5p is KLF13 in the inflammatory signaling pathway by HEK293, THP-1 and mouse macrophage line Raw264.7.Thirdly, by looking for the regulation cytokinesof miR-525-5p, such as LPS,and identify the specific sites of the cytokine activated transcription factor binding to miR-525-5p promoter, it will improve the feedback regulation loop based on miR-525-5p. In the end, it can provide clues for miR-525-5p to participate in the pathogenic mechanism in disease activity of pSLEby KLF13-RANTES pathway. It may provide meaningful biomarkers for clinical monitoring of disease activity and new treatment of pSLE.

细胞因子紊乱是pSLE病情活动的重要机制之一，而某些miRNA可以引起细胞因子表达紊乱，进而导致pSLE病情活动。我们前期研究发现miR-525-5p表达水平与pSLE病情活动度正相关，而这个过程可能通过靶基因KLF13的表达异常、进而引起RANTES等细胞因子分泌增加、最终导致pSLE病情加重。在此研究基础上，本项目拟利用病情活动前后的pSLE外周血单个核细胞、活化T细胞、293A细胞以及狼疮样小鼠模型中过表达或沉默miR-525-5p分子，从分子层面明确miR-525-5p引起RANTES等细胞因子表达水平紊乱的机制；然后通过在HEK293，THP-1以及小鼠巨噬细胞系Raw264.7中验证miR-525-5p在这个炎症信号通路中的作用靶点为KLF13基因，以及调控的分子机制；最后通过寻找调控miR-525-5p表达水平的炎症因子，如LPS，明确该炎症因子激活的转录因子结合到miR-5

细胞因子紊乱是pSLE病情活动的重要机制之一，而某些miRNA可以引起细胞因子表达紊乱，进而导致pSLE病情活动。本研究通过对新发病的3例儿童系统性红斑狼疮患儿，治疗前后血清单个核细胞miR-525-5p的测定，明确活动期外周血单个核细胞miR-525-5p水平高于稳定期。同时发现：病情活动度降低越多，外周血单个核细胞miR-525-5p水平下降越多。这为miR-525-5p用于临床监测病情活动提供了可靠依据。通过对新发病的33例儿童系统性红斑狼疮患儿血清细胞因子的测定，发现pSLE病情与细胞因子密切相关。在此基础上，通过单细胞测序技术发现pSLE外周血单核细胞比例较正常人增高、且以CD16-单核细胞为主，而后者参与细胞因子通路、发生免疫反应，进而推测：pSLE外周血单核细胞中miR-525-5p通过水平的调控THBS1基因，导致单核细胞活化，进而引起细胞因子分泌，导致pSLE病情活动。这为我们下一步研究提供了线索和依据。