中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2017, Vol. 12 ›› Issue (10): 1708-1715.doi: 10.4103/1673-5374.217351

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

miR-30c 促进周围神经损伤后许旺细胞的再髓鞘化

  

  • 收稿日期:2017-06-02 出版日期:2017-10-15 发布日期:2017-10-15
  • 基金资助:

    江苏省自然科学基金项目（BK20150409）；江苏省高校自然科学基金项目（15KJB180013）；南通市自然科学基金项目（MS12015043）；中国博士后科学基金项目（2016M600435）；江苏省博士后科学基金项目（1601056A）；江苏省高校重点学科建设项目

miR-30c promotes Schwann cell remyelination following peripheral nerve injury

Sheng Yi, Qi-hui Wang, Li-li Zhao, Jing Qin, Ya-xian Wang, Bin Yu, Song-lin Zhou   

  1. Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China
  • Received:2017-06-02 Online:2017-10-15 Published:2017-10-15
  • Contact: Song-lin Zhou, M.D.,songlin.zhou@ntu.edu.cn
  • Supported by:

    This study was supported by the Natural Science Foundation of Jiangsu Province, China, No. BK20150409; the Natural Science Foundation of Jiangsu Higher Education Institutions of China, No. 15KJB180013; the Natural Science Foundation of Nantong of Jiangsu Province, No. MS12015043; Postdoctoral Science Foundation of China, No. 2016M600435; Postdoctoral Science Foundation of Jiangsu Province of China, No. 1601056A; and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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摘要：

课题组前期研究已筛选出77个在损伤近端神经断端中差异表达的miRNAs，其中一部分miRNAs表达在周围神经损伤后先下调，随后逐渐上调，这可能与许旺细胞表型转化有关。miR-30c是其中变化最为显著的miRNA，因此实验拟进一步探讨其在坐骨神经损伤后许旺细胞再髓鞘的作用。实验设计随机选取42只成年雄性SD大鼠：36只建立坐骨神经切断模型，6只作为假手术组。分别在1, 4, 7，14，21和28d收集坐骨神经断端提取RNA。实时定量PCR显示神经损伤后miR-30c水平逐渐下降，在第4d达到最低值，随后逐渐升高。继而进行了两部分实验。体外实验：从乳鼠坐骨神经断端分离培养许旺细胞，经miR-30c激动剂转染后与背根神经节共培养, 见miR-30c的增强表达促进了与背根神经节共培养的许旺细胞中髓鞘相关蛋白的表达；体内实验：选取12只成年雄性SD大鼠，建立坐骨神经挤压伤模型。随机分为2组，在损伤后当天及7d时神经外膜下注射miR-30c激动剂或阴性对照液。损伤后14d，取坐骨神经残端组织进行免疫组化染色，Western Blot 分析及透射电镜观察。发现直接注射miR-30c可以刺激髓鞘的形成从而促进了周围神经的再生。总之，miR-30c可刺激周围神经损伤后的许旺细胞形成髓鞘，为周围神经损伤的修复提供了新的治疗靶点。

orcid:0000-0001-8598-0922(Song-lin Zhou)

Abstract:

Differential expression of miRNAs occurs in injured proximal nerve stumps and includes miRNAs that are firstly down-regulated and then gradually up-regulated following nerve injury. These miRNAs might be related to a Schwann cell phenotypic switch. miR-30c, as a member of this group, was further investigated in the current study. Sprague-Dawley rats underwent sciatic nerve transection and proximal nerve stumps were collected at 1, 4, 7, 14, 21, and 28 days post injury for analysis. Following sciatic nerve injury, miR-30c was down-regulated, reaching a minimum on day 4, and was then upregulated to normal levels. Schwann cells were isolated from neonatal rat sciatic nerve stumps, then transfected with miR-30c agomir and co-cultured in vitro with dorsal root ganglia. The enhanced expression of miR-30c robustly increased the amount of myelin-associated protein in the co-cultured dorsal root ganglia and Schwann cells. We then modeled sciatic nerve crush injury in vivo in Sprague-Dawley rats and tested the effect of perineural injection of miR-30c agomir on myelin sheath regeneration. Fourteen days after surgery, sciatic nerve stumps were harvested and subjected to immunohistochemistry, western blot analysis, and transmission electron microscopy. The direct injection of miR-30c stimulated the formation of myelin sheath, thus contributing to peripheral nerve regeneration. Overall, our findings indicate that miR-30c can promote Schwann cell myelination following peripheral nerve injury. The functional study of miR-30c will benefit the discovery of new therapeutic targets and the development of new treatment strategies for peripheral nerve regeneration.