中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (3): 671-682.doi: 10.4103/1673-5374.350214

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

内源性许旺细胞中miR-221-3p/miR-338-3p序贯表达可作为促进周围神经再生和功能恢复的策略

  

  • 出版日期:2023-03-15 发布日期:2022-08-28
  • 基金资助:

    中国国家自然科学基金项目（81771351），国家重点研发计划项目（2017YFA0105802），辽沈材料科学国家实验室联合研究基金项目（2019JH3/30100022），国家博士后科学基金项目（2018M641732）

Sequential expression of miR-221-3p and miR-338-3p in Schwann cells as a therapeutic strategy to promote nerve regeneration and functional recovery

Li-Li Wen1, Tian-Hao Yu2, Yi-Zhan Ma1, Xiao-Yan Mao1, Tian-Rang Ao3, Rabia Javed1, Hirotomo Ten4, Akira Matsuno4, Qiang Ao1, 5, *   

  1. 1Department of Tissue Engineering, School of Intelligent Medicine, China Medical University, Shenyang, Liaoning Province, China; 2The VIP Department, School and Hospital of Stomatology, Liaoning Provincial Key Laboratory of Oral Diseases, China Medical University, Shenyang, Liaoning Province, China; 3Department of Oncology, Cancer Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China; 4Department of Neurosurgery, Teikyo University School of Medicine, Tokyo, Japan; 5NMPA Key Laboratory for Quality Research and Control of Tissue Regenerative Biomaterial & Institute of Regulatory Science for Medical Device & National Engineering Research Center for Biomaterials, Sichuan University, Chengdu, Sichuan Province, China
  • Online:2023-03-15 Published:2022-08-28
  • Contact: Qiang Ao, PhD, aoqiang@tsinghua.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81771351, the National Key R&D Program of China, No. 2017YFA0105802, the Joint Research Fund Liaoning-Shenyang National Laboratory for Materials Science, No. 2019JH3/30100022, and the National Science Foundation for Post-doctoral Scientists of China, No. 2018M641732 (all to QA and LLW).

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摘要：

内源性许旺细胞在神经修复过程中的功能特性是动态的，优化自体许旺细胞在神经修复不同阶段的功能十分重要。既往有研究显示，miR-221-3p可促进许旺细胞的增殖和迁移，而miR-338-3p能促进许旺细胞髓鞘的形成。实验首先以导管桥接横断坐骨神经构建神经再生室大鼠模型，并将携带miRNA-221常规慢病毒载体和强力霉素诱导的携带miRNA-338-Tet on的慢病毒载体系统共同注射于神经断端导管腔内来有序调控内源性许旺细胞不同阶段的生物学功能。结果显示：（1）可以有序调节许旺细胞的生物学功能，增加有髓轴突的直径和密度以及髓鞘中NF200和MBP的蛋白表达，从而改善损伤周围神经的功能。（2）通过miRDB数据库预测、nanopore全转录组基因测序、定量PCR和双荧光素酶实验检测发现，Cdkn1b和Nrp1分别被预测和验证为miR-221-3p和miR-338-3p的靶基因，且体外实验证实了其对许旺细胞的调节作用。（3）该实验创新性地通过有序调节许旺细胞生物学特性来促进周围神经再生，为周围神经损伤的治疗建立了新的概念和模型，会对临床上治疗坐骨神经损伤产生直接的指导意义。

https://orcid.org/0000-0002-1309-1971 (Qiang Ao)

关键词:

周围神经损伤, 神经再生, 序贯表达, 微小RNA, 调控, 内源性许旺细胞, miR-221, miR-338, cdkn1b, Nrp1

Abstract: The functional properties of endogenous Schwann cells (SCs) during nerve repair are dynamic. Optimizing the functional properties of SCs at different stages of nerve repair may have therapeutic benefit in improving the repair of damaged nerves. Previous studies showed that miR-221-3p promotes the proliferation and migration of SCs, and miR-338-3p promotes the myelination of SCs. In this study, we established rat models of sciatic nerve injury by bridging the transected sciatic nerve with a silicone tube. We injected a miR-221 lentiviral vector system together with a doxycycline-inducible Tet-On miR-338 lentiviral vector system into the cavity of nerve conduits of nerve stumps to sequentially regulate the biological function of endogenous SCs at different stages of nerve regeneration. We found that the biological function of SCs was sequentially regulated, the diameter and density of myelinated axons were increased, the expression levels of NF200 and myelin basic protein were increased, and the function of injured peripheral nerve was improved using this system. miRNA Target Prediction Database prediction, Nanopore whole transcriptome sequencing, quantitative PCR, and dual luciferase reporter gene assay results predicted and verified Cdkn1b and Nrp1 as target genes of miR-221-3p and miR-338-3p, respectively, and their regulatory effects on SCs were confirmed in vitro. In conclusion, here we established a new method to enhance nerve regeneration through sequential regulation of biological functions of endogenous SCs, which establishes a new concept and model for the treatment of peripheral nerve injury. The findings from this study will provide direct guiding significance for clinical treatment of sciatic nerve injury.

Key words: cdkn1b, miR-221, miR-338, miRNA, nerve regeneration, Nrp1, peripheral nerve injury, regulation, Schwann cells, sequential expression