中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (2): 265-271.doi: 10.4103/1673-5374.226397

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

差异miRNA及靶基因在缺血再灌注损伤及apelin-13干预过程中发挥重要作用的高通量测序分析

  

  • 收稿日期:2017-10-23 出版日期:2018-02-15 发布日期:2018-02-15
  • 基金资助:

    中国国家自然科学基金项目（81501018，816712276），山东省自然科学基金项目（ZR2013CQ031，ZR2014HL040）

High-throughput sequencing analysis of differentially expressed miRNAs and target genes in ischemia/reperfusion injury and apelin-13 neuroprotection

Chun-mei Wang, Xue-lu Yang, Ming-hui Liu, Bao-hua Cheng, Jing Chen, Bo Bai   

  1. Neurobiology Institute, Jining Medical University, Jining, Shandong Province, China
  • Received:2017-10-23 Online:2018-02-15 Published:2018-02-15
  • Contact: Jing Chen, Ph.D. or Bo Bai,Ph.D., chenjnmc@163.com or bobai@mail.jnmc.edu.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81501018 and 816712276; the Natural Science Foundation of Shandong Province of China, No. ZR2013CQ031 and ZR2014HL040.

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摘要：

miRNA可通过与靶mRNA3’端非编码区配对调控基因表达，从而调节多种生物学过程，研究发现miRNA在脑缺血再灌注损伤模型中表达异常。高通量测序是近年发展起来的一种基于合成技术测序识别miRNA的方法，已广泛用于生物miRNA的分析。因此实验对再灌注前侧脑室注射50 ng/kg apelin-13的脑缺血再灌注损伤模型大鼠进行分析。先以高通量测序分析脑缺血再灌注模型大鼠大脑海马组织中miRNA表达谱变化，见352个miRNA表达存在差异，其中78个miRNA的表达差异大于1倍。然后以实时荧光定量PCR对其中4个表达水平发生明显变化的miRNA进行验证，得到的结果与高通量测序分析结果一致。基因本体分析结果显示，差异基因参与细胞、代谢过程、单生物代谢、细胞骨钉等生物过程。KEGG分析显示，差异基因参与代谢、MAPK、钙离子以及NF-κB信号通路。结果说明差异miRNA及靶基因在模型大鼠缺血再灌注损伤及apelin-13干预过程中发挥了重要的作用。

orcid:0000-0002-7198-2393(Jing Chen)
         0000-0001-9988-9318 (Bo Bai)

关键词: 神经再生, RNA测序, microRNA, apelin, 缺血再灌注损伤, 神经保护, 神经肽, 高通量测序分析

Abstract:

miRNAs regulate a variety of biological processes through pairing-based regulation of gene expression at the 3′ end of the noncoding region of the target miRNA. miRNAs were found to be abnormally expressed in ischemia/reperfusion injury models. High-throughput sequencing is a recently developed method for sequencing miRNAs and has been widely used in the analysis of miRNAs. In this study,ischemia/reperfusion injury models were intracerebroventricularly injected with 50 μg/kg apelin-13. High-throughput sequencing showed that 357 known miRNAs were differentially expressed among rat models, among which 78 changed to > 2-fold or < 0.5-fold. Quantitative real-time polymerase chain reaction was selected to confirm the expression levels of four miRNAs that were differentially expressed,the results of which were consistent with the results of high-throughput sequencing. Gene Ontology analysis revealed that the predicted targets of the different miRNAs are particularly associated with cellular process, metabolic process, single-organism process, cell, and binding. Kyoto Encyclopedia of Gene and Genome analysis showed that the target genes are involved in metabolic pathways, mitogen-activated protein kinase signaling pathway, calcium signaling pathway, and nuclear factor-κB signaling pathway. Our findings suggest that differentially expressed miRNAs and their target genes play an important role in ischemia/reperfusion injury and neuroprotection by apelin-13.

Key words: nerve regeneration, RNA sequencing, microRNA, apelin, ischemia/reperfusion injury, neuroprotection, neuropeptide, high-throughput sequencing, neural regeneration