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 (7): 1253-1262.doi: 10.4103/1673-5374.235073

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

周围神经再生过程中的时空microRNA谱：miR-138靶向波形蛋白并抑制许旺细胞迁移和增殖

  

  • 收稿日期:2018-05-16 出版日期:2018-07-15 发布日期:2018-07-15
  • 基金资助:

    美国Leisa V. Clayton基金资助

Spatiotemporal microRNA profile in peripheral nerve regeneration: miR-138 targets vimentin and inhibits Schwann cell migration and proliferation

Travis B. Sullivan1, Litchfield C. Robert2, Patrick A. Teebagy1, Shannon E. Morgan1, Evan W. Beatty1, Bryan J. Cicuto3, Peter K. Nowd1, Kimberly M. Rieger-Christ1, David J. Bryan2, 3   

  1. 1 Department of Translational Research, Lahey Hospital & Medical Center, Burlington, MA, USA
    2 Tissue Engineering Laboratory, Lahey Hospital & Medical Center, Burlington, MA, USA
    3 Department of Plastic and Reconstructive Surgery, Lahey Hospital & Medical Center, Burlington, MA, USA
  • Received:2018-05-16 Online:2018-07-15 Published:2018-07-15
  • Contact: Kimberly Rieger-Christ, Ph.D.,kimberly.r.christ@lahey.org.
  • Supported by:

    This study was supported by the Leisa V. Clayton Foundation.

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摘要：

波形蛋白位于轴突生长前端，并与神经再生活动相关，比如髓鞘形成、神经可塑性、神经轴质中的激酶信号传导和细胞迁移，但调控许旺细胞内波形蛋白表达的相关机制尚未探索。为剖析大鼠坐骨神经横断后再生过程中微小RNA（miRNA）的空间和时间表达谱，并探讨miR-138-5p靶向波形蛋白在SC增殖和迁移中的潜在作用。实验使用聚乙烯神经导管桥接大鼠坐骨神经两断端，检测神经再生期间7,14,30,60和90天的miRNA表达。使用微阵列分析确定miRNA表达的相对水平，并随后用定量实时聚合酶链反应进行验证。评估miRNA模拟物转染的许旺细胞迁移和增殖能力。发现7种失调miRNA涉及细胞迁移，GO和KEGG分析预测了伤口愈合所必需基因的表达活性。将其中的miRNA-138-5p转染到许旺细胞中，发现细胞迁移和增殖受到抑制。miR-138-5p在癌细胞中直接靶向波形蛋白，并且在大鼠许旺细胞中进行的荧光素酶测定证实了这一点。这些结果详细描述了miR-138-5p在大鼠外周神经再生中的作用，体现了其在周围神经系统再生方面的重要调节作用。

orcid:0000-0002-8425-132X(Kimberly Rieger-Christ)

关键词: 非编码RNA, 神经再生, 神经导向, 坐骨神经切断, 周围神经损伤, 伤口愈合, 基因本体过程, 京都基因和基因组通路百科全书, 芯片, 萤光素酶测定

Abstract:

While the peripheral nervous system has regenerative ability, restoration of sufficient function remains a challenge. Vimentin has been shown to be localized in axonal growth fronts and associated with nerve regeneration,including myelination, neuroplasticity, kinase signaling in nerve axoplasm, and cell migration;however, the mechanisms regulating its expression within Schwann cell (SC) remain unexplored. The aim of this study was to profile the spatial and temporal expression profile of microRNA (miRNA) in a regenerating rat sciatic nerve after transection, and explore the potential role of miR-138-5p targeting vimentin in SC proliferation and migration. A rat sciatic nerve transection model, utilizing a polyethylene nerve guide,was used to investigate miRNA expression at 7, 14, 30, 60, and 90 days during nerve regeneration. Relative levels of miRNA expression were determined using microarray analysis and subsequently validated with quantitative real-time polymerase chain reaction. In vitro assays were conducted with cultured Schwann cells transfected with miRNA mimics and assessed for migratory and proliferative potential. The top seven dysregulated miRNAs reported in this study have been implicated in cell migration elsewhere, and GO and KEGG analyses predicted activities essential to wound healing. Transfection of one of these, miRNA-138-5p, into SCs reduced cell migration and proliferation. miR-138-5p has been shown to directly target vimentin in cancer cells, and the luciferase assay performed here in rat Schwann cells confirmed it. These results detail a role of miR-138-5p in rat peripheral nerve regeneration and expand on reports of it as an important regulator in the peripheral nervous system.

Key words: non-coding RNA, neural regeneration, nerve guide, sciatic nerve transection, peripheral nerve injury, wound healing, Gene Ontology processes, Kyoto Encyclopedia of Genes and Genomes pathways, microarray, luciferase assay