系统性红斑狼疮患者外周血B淋巴细胞中狼疮活动相关miRNA的筛选  

Screening of the miRNAs related to disease activity in peripheral blood B lymphocytes of systemic lupus erythematosus patients

作　　者：陈芳[1] 陈朝生[1] 章慧娣[1] 薛向阳[2] 丁晓凯[1] 孙莉[3] 陈丹[3] 黄朝兴[1] 

机构地区：[1]温州医学院附属第一医院肾内科,浙江温州325000 [2]温州医学院微生物学与免疫学教研室,浙江温州325000 [3]温州医学院附属第一医院风湿科,浙江温州325000

出　　处：《温州医学院学报》2013年第2期106-111,共6页Journal of Wenzhou Medical College

基　　金：温州市科技局科研基金资助项目(Y20100198)

摘　　要：目的:通过筛选系统性红斑狼疮(SLE)患者外周血B淋巴细胞中与狼疮活动相关的microRNA(miRNA),探讨B淋巴细胞参与SLE发病的机制。方法:选取45例重度活动期SLE患者作为研究对象,另选20例稳定期SLE患者和10例健康体检者分别作为对照,采集外周静脉血,用磁珠阴性分选法分离B淋巴细胞并提取RNA,将每组单个样本RNA混成组总RNA,进行组间芯片分析。实时荧光定量PCR(RealtimePCR)验证miRNA芯片检测结果。结果:重度活动期SLE患者外周血B淋巴细胞中有16条差异表达的miRNA,其中下调10条,上调6条。Realtime PCR对其中2条(miR-181a、miR-103)miRNA的验证结果与芯片检测所示具有较好的一致性。结论:SLE患者病情活动性与B淋巴细胞中miRNA异常表达密切相关。Objective： To explore the pathogenesis of B lymphocytes participated in SLE through thescreening of the microRNAs （miRNAs） related to lupus activity in peripheral blood B lymphocytes of systemic lupus erythmatosus （SLE） patients. Methods： Forty-five cases of severe active SLE patients as the researchobject, 20 cases of remission SLE patients and 15 healthy controls as control groups were selected respectively. The peripheral EDTA anticoagulation blood was obtained to extract miRNAs in B lymphocytes separated bymagnetic cell sorting system, mix all of the RNA of B lymphocytes in the same group, and compared the miRNAs profile of these 3 groups one to one by the miRNA microarray chip analysis. The microarray results were validatedby realtmie PCR. Results： The miRNA microarray chip analysis identified 16 miRNAs differently expressed in severe active SLE patients, among which 10 down-regulated and 6 up-regulated. The findings of realtmie PCR on2 miRNA （miR-181a, miR-103） were in high concordance with microarray results. Conclusion： The disease activity of SLE is related to the abnormal expression of miRNAs in peripheral blood B lymphocytes.

关 键 词：红斑狼疮 系统性 MIRNA 免疫磁珠 B淋巴细胞 芯片 

分 类 号：R593.24[医药卫生—内科学]