基础医学与临床 ›› 2016, Vol. 36 ›› Issue (6): 739-746.

• 研究论文 • 上一篇    下一篇

COPD动态miRNA表达谱的构建及生物信息学分析

谢丽华1,孙圣华1,卢俊娟2,林桦1   

  1. 1. 中南大学湘雅三医院
    2. 湘雅三医院
  • 收稿日期:2015-11-30 修回日期:2016-03-25 出版日期:2016-06-05 发布日期:2016-05-27
  • 通讯作者: 孙圣华 E-mail:sunshenghua03@163.com
  • 基金资助:
    国家自然科学基金

Construction of dynamic miRNA expression profiling and bioinformatics analysis in COPD

  • Received:2015-11-30 Revised:2016-03-25 Online:2016-06-05 Published:2016-05-27
  • Supported by:
    the National Natural Science Foundation of China

摘要: 目的 构建慢性阻塞性肺疾病(COPD)动态大鼠模型不同时期肺组织miRNA表达谱,并对差异表达的miRNAs进行生物信息学分析,以了解COPD发生发展中miRNAs变化规律及可能涉及相关机制。方法 1)将大鼠随机分为对照组、吸烟2、4、6、8周组和15周+气管内滴注猪胰弹性蛋白酶组,每组5只,病理学观察其形态学改变;2)取对照组、吸烟4周和15周+气管内滴注猪胰弹性蛋白酶组大鼠肺组织各2例,用于miRNA表达谱芯片分析;3)实时荧光定量PCR法验证芯片数据;4)运用生物信息学法对miRNA表达谱进行靶基因预测及聚类分析。结果 1)成功建立COPD动态大鼠模型,该鼠肺组织早期以出现炎症细胞浸润为主,晚期出现肺气肿改变的动态病理变化;2)成功建立miRNAs动态表达谱,熏烟4周组共30个miRNAs异常表达,熏烟15周组共37个miRNAs异常表达;3)实时荧光定量PCR法证实miR-146a、 miR-21、 miR-206和 miR-181a变化趋势与芯片一致;4)生物信息学分析显示异常表达的miRNAs可能与COPD发病机制密切相关。结论 1)成功构建动态COPD大鼠模型及miRNA表达谱。2)通过生物信息学分析异常表达的miRNA可能在COPD发病机制中发挥重要作用。

关键词: 慢性阻塞性肺疾病, miRNA, 微芯片, 生物信息学

Abstract: Objective The miRNA expression profiling in different lung tissue of COPD dynamic rat model was established. The differential expression miRNAs were analysed by bioinformatics methods.This study is aimed at finding the variety rule of miRNAs and possibly refer to the correlate pathogenesis of COPD. Methods 1 The rats were randomly divided into the control group; the cigarette smoking (CS) for 2、4、6、8w group and 15w plus elastase-treated group. Each group had five rats. Morphological changes of the lungs were performed through pathology observation. 2 The lung tissue of control group、CS for 4w group and 15w plus elastase-treated group(each group had two rats)were analysed by miRNA microarray. 3 The dysregulated miRNAs were detected by qRT-PCR for the further confirming of microarray data. 4 The miRNA expression profiling was analysed by bioinformatics methods including target prediction and unsupervised hierarchical clustering analysis.Results 1 COPD dynamic rat model were established successfully. The dynamic pathology changes in the lung tissue of this model were inflammatory infiltrates in early stage and emphysema in late stage. 2 The dynamic miRNA expression profiling was established. There were 30 and 37 differentially expressing miRNAs in the lungs of rats with 4w group and 15w group respectively. 3 Further qRT-PCR indicated that the relative expression levels of miR-146a, miR-21, miR-206 and miR-181a were consistent with the microarray data. 4 The differential expression miRNAs may be closely related to the pathogenesis of COPD through bioinformatics analysis. Conclusions:1 The dynamic rats models of COPD and miRNA expression profiling were established successfully. 2 The differential expression miRNAs may play important role in the pathogenesis of COPD through bioinformatics analysis.

Key words: Chronic obstructive pulmonary disease (COPD), miRNA, microarray, Bioinformatics

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