缺血性卒中不同亚型患者血浆miRNAs表达谱变化分析  

Analysis of plasma miRNAs expression profile in different subtypes of ischemic stroke

作　　者：仲倩维 马爱军[1] 潘旭东[1] 杨淑娜[1] 王兰[1] 张璋[1] 庞萌[1] 

机构地区：[1]青岛大学附属医院神经内科,266000

出　　处：《中华神经科杂志》2015年第2期114-119,共6页Chinese Journal of Neurology

基　　金：山东省自然科学基金资助项目（ZR2013HM022）

摘　　要：目的研究缺血性卒中不同亚型患者血浆中miRNAs表达谱的差异。方法选取2012年11—12月于青岛大学附属医院神经内科就诊的首次发病的16例急性缺血性卒中患者，按TOAST分型将其分为大动脉粥样硬化性（large artery atherosclerotic，LAA）卒中组8例，小动脉闭塞性（small artery occlusion，SAO）卒中组8例，以及同期健康体检者8名作为对照组，应用高通量测序法检测各组血浆中miRNAs的表达谱，筛选出差异表达的miRNAs，实时荧光定量PCR验证结果，并进行靶基因预测及生物信息学分析。结果LAA组、SAO组及对照组血浆中分别检测到369个miRNAs成熟体。通过miRNAs差异分析，发现let-7a-5p、miR-744—5p等12个miRNAs在3组中均呈现差异性表达（表达差异均大于2倍，P〈0．01）；与对照组比较，miR-126-5p、miR-23a-3p、miR-143—3p等34个miRNAs在LAA组表达显著下调（表达差异均大于2倍，P〈0．01）；在SAO组中miR-1304-3p、miR-451a较对照组表达显著下调（表达差异均大于2倍，P〈0．01），而miR-320a、miR-320b、miR-99b-5p等27个miRNAs较对照组表达则显著上调（表达差异均大于2倍，P〈0．01）。实时荧光定量PCR对其中的miR-146b-5p、miR-23a-3p、miR-451a的验证结果与高通量检测结果一致。生物信息学分析发现，差异表达的miRNAs调控的靶基因主要与细胞增殖、黏附、代谢等生物学功能相关。结论LAA型卒中患者及SAO型卒中患者血浆miRNAs表达谱存在明显差异，提示miRNAs可能通过靶基因在LAA及SAO的发病过程中发挥不同的调节作用。Objective To investigate the changes of circulating miRNAs expression profiles in different subtypes of ischemic stroke according to the Trial of Org 10 172 in Acute Stroke Treatment. Methods We selected 16 patients diagnosed as acute ischemic stroke at first time in the Department of Neurology, the Affiliated Hospital of Medical College Qingdao University from November to December in 2012. They were divided into large artery atherosclerosis （LAA） stroke group （n = 8） and small artery occlusive （SAO） stroke group （ n = 8 ）. At the same time, 8 healthy checkup subjects were selected as control. High-throughput sequencing was used to detect the expression profiles of miRNAs in the plasma, and the high-throughput sequencing results were validated by quantitative real-time polymerase chain reaction. We performed the miRNAs variance analysis and associated bioinformatics analysis. Results Thirty hundred and sixty-nine miRNAs were detected in LAA group, SAO group and the control group. We found remarkable differences （ fold change 〉 2, P 〈 0. 01 ） in the expression of 12 miRNAs, including let-7a-5p, miR-744-Sp, etc. , between any two groups of the three groups. Thirty-four miRNAs, containing miR-126- 5p, miR-23a-3p, miR-143-3p, etc. , had a lower expression （fold change 〉 2, P 〈 0. 01 ） in LAA group than that in the control group. In comparison with the control group, miR-1304-3p and miR-451a were significantly down-regulated （ fold change 〉 2, P 〈 0. 01 ） , while 27 miRNAs were significantly up-regulated （fold change 〉2, P 〈0. 01 ） in SAO group. The expression levels of miR-146b-5p, miR-23a-3p and miR451a were validated in accordance with the results of real-time PCR. Target gene prediction and functional analysis revealed that target genes regulated by differential expression of miRNAs were mainly associated with cell proliferation, adhesion, metabolism and other biological functions. Conclusion miRNAs are differently expressed in the plasma of LAA group, SAO group and

关 键 词：脑缺血 卒中 微RNAS 高通量筛选分析 动脉粥样硬化 

分 类 号：R743.3[医药卫生—神经病学与精神病学]