无症状性脑动脉粥样硬化与大动脉粥样硬化性卒中患者血浆微小RNA表达分析

摘要

Objective To establish the plasma miRNAs expression profile change of patients with asymptomatic cerebral atherosclerotic (AS) and large artery atherosclerotic (LAA) ischemic stroke. Methods Retrospective study is adopted, and our study included eight asymptomatic cerebral atherosclerotic patients (AS group), eight LAA ischemic stroke patients (LAA group), and eight controls (control group). All participants were selected from the Affiliated Hospital of Medical College Qingdao University from January 1 to Februry 28 in 2013. The Solexa sequencing technology was employed to detect the expression proifle of miRNAs in the specimens, we found out the differentially expressed miRNAs. At last, we made an initial prediction and functional analysis of target genes regulated by the differentially expressed miRNAs. The results of the Solexa sequencing technology were veriifed by real-time polymerase chain reaction (PCR). Results Twenty six miRNAs including miR-let-7a-5p, miR-146b-5p, miR-26a-5p, miR-23a-3p were found differentially expressed among the 3 groups. Compared with control group, 41 miRNAs including miR-10b-5p, miR-126-5p, miR-143-3p, miR-192-5p were found differentially expressed in AS group and LAA group. The bioinformatics analysis indicated that the target genes of these differentially expressed miRNAs participated in cell adhesion, differentiation, and proliferation. The results of real-time PCR indicated that there was a good consistency between PCR veriifed results and Solexa sequencing data. Conclusion miRNA expression proifle of patients with AS and LAA ischemic stroke has some similarities, also has some differences.%目的建立无症状性脑动脉粥样硬化（atherosclerosis，AS）及大动脉粥样硬化（large artery atherosclerotic，LAA）性卒中患者血浆微小核糖核苷酸（microribonucleic acid，miRNA）差异表达谱。　　方法收集2013年1月～2013年2月于青岛大学医学院附属医院神经内科就诊的8名AS患者（AS组）、LAA性卒中患者（LAA组）和对照者（control组）的空腹前臂静脉血血浆标本，应用Solexa高通量测序技术检测每组血浆标本miRNA表达谱，筛选出差异表达的miRNA，并进行靶基因预测和功能分析，采用实时荧光定量聚合酶链反应（polymerase chain reaction，PCR）技术对高通量检测结果进行验证。　　结果3组间比较，表达差异均具有显著性的miRNA有miR-let-7a-5p、miR-146b-5p、miR-26a-5p、miR-23a-3p等26个；在AS组与LAA组表达一致，较对照组表达差异具有显著性的miRNA有miR-10b-5p、miR-126-5p、miR-143-3p、miR-192-5p等41个。本研究生物信息学分析发现，差异miRNAs调控的靶基因主要与细胞黏附、分化、增殖等生物学过程相关。实时荧光定量PCR检测miR-146b-5p、miR-23a-3p、miR-10b-5p，结果与高通量检测结果一致。　　结论 AS患者及LAA性卒中患者血浆miRNAs表达谱既有差别又有相同之处。