中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (9): 1865-1870.doi: 10.4103/1673-5374.306092

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

转录组学分析发现周围神经损伤后必需的microRNAs

  

  • 出版日期:2021-09-15 发布日期:2021-02-05
  • 基金资助:

    本研究得到国家自然科学基金(31971276);江苏省高等学校自然科学基金(19KJA32005)

Transcriptomic analysis reveals essential microRNAs after peripheral nerve injury

Yu Wang, Shu Wang, Jiang-Hong He*   

  1. Key Laboratory for Neuroregeneration of Jiangsu Province and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China
  • Online:2021-09-15 Published:2021-02-05
  • Contact: Jiang-Hong He, PhD, hejh@ntu.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 31971276 (to JHH) and the Natural Science Foundation of Jiangsu Higher Education Institutions of China (Major Program), No. 19KJA320005 (to JHH). 

摘要:

一些研究证实,MicroRNAs (miRNAs)可介导靶基因转录后的调控,并参与周围神经损伤等多种生理和病理过程,但其参与和表达等生物学模式尚不明确。课题组在前期实验中检测了不同阶段坐骨神经损伤(急性期、亚急性期和急性后期)后的基因变化,并初步得到了大鼠坐骨神经损伤后的基因表达谱。在此基础上,本次实验拟联合分析大鼠坐骨神经损伤后不同时期的mRNA和miRNAs的具体表达变化,并使用生物信息学软件Pathway Analysis (IPA)研究差异表达mRNA的上游miRNAs的参与模式。①实验发现大鼠坐骨神经夹伤后的1,4,7,14 d,在神经损伤残端分别检测出31,42,30,23个相应上游miRNAs(如miR-21,let-7,miR-223,miR-10b,miR-132,miR-15b,miR-127,miR-29a,miR-29b和miR-9)②实验还进一步分析了这些miRNAs的差异表达模式发现,miR-21,miR-132,miR-29a,miR-29b等miRNAs在坐骨神经损伤后表达均上调,这些miRNAs涉及的生物学过程主要分为多细胞组织对应激的反应、表皮生长因子受体信号通路的正调控、上皮细胞分化的负调控和心肌组织生长的调节等;③实验还成功构建了miRNAs中作用差异表达最显著的let-7、miR-21和miR-223的调控网络;④上述结果证实,在大鼠坐骨神经损伤后损伤神经残端可发现let-7、miR-21和miR-223等多个差异表达的miRNAs,这些miRNAs在周围神经再生中可能发挥重要的调节作用。实验已于2019年3月3日获得江苏省实验动物伦理委员会批准(批准号:20190303-18)。

https://orcid.org/0000-0002-5185-7982 (Jiang-Hong He)

关键词:

周围神经损伤, 坐骨神经夹伤, RNA测序, 生物信息学分析, IPA, microRNA, 调控网络, 周围神经再生

Abstract: Studies have shown that microRNAs (miRNAs) mediate posttranscriptional regulation of target genes and participate in various physiological and pathological processes, including peripheral nerve injury. However, it is hard to select key miRNAs with essential biological functions among a large number of differentially expressed miRNAs. Previously, we collected injured sciatic nerve stumps at multiple time points after nerve crush injury, examined gene changes at different stages (acute, sub-acute, and post-acute), and obtained mRNA expression profiles. Here, we jointly analyzed mRNAs and miRNAs, and investigated upstream miRNAs of differentially expressed mRNAs using Ingenuity Pathway Analysis bioinformatic software. A total of 31, 42, 30, and 23 upstream miRNAs were identified at 1, 4, 7, and 14 days after rat sciatic nerve injury, respectively. Temporal expression patterns and biological involvement of commonly involved upstream miRNAs (miR-21, let-7, miR-223, miR-10b, miR-132, miR-15b, miR-127, miR-29a, miR-29b, and miR-9) were then determined at multiple time points. Expression levels of miR-21, miR-132, miR-29a, and miR-29b were robustly increased after sciatic nerve injury. Biological processes involving these miRNAs include multicellular organismal response to stress, positive regulation of the epidermal growth factor receptor signaling pathway, negative regulation of epithelial cell differentiation, and regulation of myocardial tissue growth. Moreover, we constructed mechanistic networks of let-7, miR-21, and miR-223, the most significantly involved upstream miRNAs. Our findings reveal that multiple upstream miRNAs (i.e., let-7, miR-21, and miR-223) were associated with gene expression changes in rat sciatic nerve stumps after nerve injury, and these miRNAs play an important role in peripheral nerve regeneration. This study was approved by the Experimental Animal Ethics Committee of Jiangsu Province of China (approval No. 20190303-18) on March 3, 2019.

Key words: bioinformatic analysis, Ingenuity Pathway Analysis, mechanistic network, microRNA, peripheral nerve injury, peripheral nerve regeneration, RNA sequencing, sciatic nerve crush