【作者】 王海燕；

【导师】 张烜； 陈梅红；

【作者基本信息】 中国协和医科大学 ， 风湿免疫， 2010， 硕士

【摘要】 目的：系统性红斑狼疮(Systemic Lupus Erythematosus, SLE)是自身免疫介导的,以免疫性炎症为突出表现的弥漫性结缔组织病,其发病机制尚不完全清楚。microRNA是一类分布广泛的小的非编码蛋白质的RNAs,在生物体内负调控基因表达。本研究旨在利用生物信息学方法和分子生物学方法,寻找出与SLE发病可能相关的microRNA,并初步探讨其在疾病中的作用。方法：通过在SLE发病中起到明确作用的编码区基因,借助于哺乳动物microRNA靶基因数据库miRanda和TargetScan,利用生物信息学分析方法查找出与SLE发病机制可能相关的microRNA。应用实时荧光定量聚合酶链反应(Real-timePCR)技术验证has-miR-146a、has-miR-181d、has-miR-7在SLE患者及正常人外周血单个核细胞中的表达差异,同时对has-miR-146a、has-miR-181d、has-miR-7在基因组中的位置及其可能作用的与SLE发病相关的靶基因进行了确定和预测,为其在疾病中的作用及其靶基因测定和参与发病的机制提供了一定的线索。结果：RT-PCR结果显示SLE患者外周血PBMCs中hsa-miR-146a的ΔCt高于正常对照组(4.52±1.18 vs.2.76±1.38), mRNA的表达量显著低于正常对照组,且两者之间有显著性统计学差异(P=0.02); hsa-miR-181d的ΔCt高于正常对照组(6.39±0.31 vs. 5.75±0.88)相比,两者之间并无统计学差异(P>0.05)；而hsa-miR-7的mRNA表达量(9.25±0.91 vs.9.30±1.08)与正常对照组相比无统计学差异(P>0.05)。结论：在SLE的发病中,microRNA可能起到了一定的作用。与正常对照相比,SLE患者PBMCs中hsa-miR-146a的表达水平降低,提示：hsa-miR-146a可能通过作用于与SLE发病相关的靶基因而参与疾病的发生。更多还原

【Abstract】 Objective. Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease, to highlight the performance of immune inflammation, diffuse connective tissue disease. Its mechanism is not entirely clear.MicroRNAs (miRNAs) have been recently identified as regulators that modulate target gene expression and are widespread class of small non-protein coding RNAs. This study aims to use bioinformatics methods and biological methods, to find out the pathogenesis of SLE may be associated with the microRNA, and to investigate its role in disease.Methods. To determine the expression spectrum of microRNAs from patients with systemic lupus erythematosus (SLE), we investigate the relationship between microRNA expressions and SLE mechanism. With the bioinformatics, we search the gene expressions related with SLE patients;with the targetgene database,we predict the microRNAs related with SLE patients.Through the encoding gene related with the SLE disease,by means of mammalian microRNA target gene database:miRanda and TargetScan,we use the biological information method to credit the microRNAs related with SLE pathogenesis. Application of real-time fluorescence quantitative polymerase chain reaction(Real-Time PCR) technology,we assayed the expressions of hsa-miR-146a、hsa-miR-181d、hsa-miR-7 in SLE patients and healthy controls. Meanwhile,the has-miR-146a, has-miR-181d, has-miR-7 position in the genome and its possible role in the target genes associated with the pathogenesis of SLE were identified and predicted, to provide some clues for its role involved in disease mechanisms.Results. Real-time PCR showed results that hsa-miR-146a mRNA was obviously decreased in PBMCs of SLE patients compared to healthy controls (ΔCt 4.52±1.18 vs. 2.76±1.38,P=0.02), but the expression of hsa-miR-181d in SLE patients was no significant difference compared to that in healthy controls(ΔCt 6.39±0.31 vs. 5.75±0.88,P>0.05), there was no significant difference in hsa-miR-7 expression between SLE patients and healthy controls(ΔCt 9.25±0.91 vs.9.30±1.08, P>0.05).Conclusion. microRNA may play a role in the pathogenesis of SLE.Compared to healthy controls, hsa-miR-146a expressions are reduced in PBMCs from patients with SLE. Thus, by acting on the target gene with SLE disease, hsa-miR-146a may be participated in disease mechanisms.更多还原