中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2025, Vol. 20 ›› Issue (1): 277-290.doi: 10.4103/1673-5374.390956

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

miRNA-21-5p是缺氧预处理骨髓神经嵴细胞促进损伤周围神经再生的重要贡献者

  

  • 出版日期:2025-01-15 发布日期:1900-01-01
  • 基金资助:
    国家自然科学基金项目（31870977）；国家重点技术研发计划项目（2017YFA0104700）；2022年度江苏省优秀博士后人才资助项目；江苏省高等学校重点学科建设〔PAPD〕，江苏省高等院校基础科学（自然科学）研究重大项目（22KJA180001）

miRNA-21-5p is an important contributor to the promotion of injured peripheral nerve regeneration using hypoxia-pretreated bone marrow–derived neural crest cells

Meng Cong1, 2, #, Jing-Jing Hu1, 3, #, Yan Yu1, Xiao-Li Li1, 4, Xiao-Ting Sun1, Li-Ting Wang1, Xia Wu2, Ling-Jie Zhu1, Xiao-Jia Yang1, Qian-Ru He2, Fei Ding2, 5, *, Hai-Yan Shi1, 2, *   

  1. 1School of Medicine, Nantong University, Nantong, Jiangsu Province, China; 2Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education and Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China; 3Department of Physiology, Jiangsu Health Vocational College, Nanjing, Jiangsu Province, China; 4Department of Pathology, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China; 5Jiangsu Clinical Medicine Center of Tissue Engineering and Nerve Injury Repair, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China
  • Online:2025-01-15 Published:1900-01-01
  • Contact: Hai-Yan Shi, PhD, haiyansh@ntu.edu.cn; Fei Ding, MS, dingfei@ntu.edu.cn.
  • Supported by:
    The study was supported by the National Natural Science Foundation of China, No. 31870977 (to HYS); the National Key Technologies Research and Development Program of China, No. 2017YFA0104700 (to FD); 2022 Jiangsu Funding Program for Excellent Postdoctoral Talent (to MC); Priority Academic Program Development of Jiangsu Higher Education Institutions [PAPD] and the Major Project of Basic Science (Natural Science) Research in Higher Education Institutions of Jiangsu Province, No. 22KJA180001 (to QRH).

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摘要：

作者既往研究已揭示作为许旺细胞前体的大鼠骨髓来源的神经嵴细胞对长距离神经缺损有修复作用。细胞治疗可通过旁分泌生物活性因子和细胞间通讯增强周围神经的修复和再生。然而，分泌组中可溶性细胞因子和细胞外囊泡内含物的贡献复杂性仍然难以捉摸。为揭示分泌组和细胞外囊泡在修复损伤周围神经的作用，实验收集缺氧预处理的神经嵴细胞条件培养基，发现其对氧糖剥夺损伤的感觉神经元具有显著的修复效果，且缺氧预处理的神经嵴细胞中多种营养因子mRNA呈高表达。其次对神经嵴细胞外囊泡进行RNA测序和生物信息学分析，可见miR-21-5p在缺氧预处理的神经嵴细胞外囊泡中富集。随后，为进一步明确富含miR-21-5p的缺氧预处理神经嵴细胞外囊泡在感觉神经元轴突生长和再生中的作用，在体外感觉神经元的微流控轴突离断模型中发现缺氧预处理的神经嵴细胞外囊泡可促进感觉神经元轴突生长与再生，且其作用明显依赖于所负载的miR-21-5p。最后，构建了负载miR-21-5p的神经导管以修复大鼠的坐骨神经缺损，发现损伤大鼠的后肢运动和感觉功能以及后肢肌肉组织形态均得到明显恢复。提示缺氧预处理神经嵴细胞外囊泡是一种高度富集miRNA-21-5p的天然纳米颗粒，且miRNA-21-5p是缺氧预处理神经嵴细胞分泌组促进神经再生的主要贡献者之一。这有助于解释分泌组和细胞外囊泡在修复损伤周围神经的作用机制，也将促进miR-21-5p在组织工程再生医学中的应用。

https://orcid.org/0000-0002-9615-1270 (Hai-Yan Shi); https://orcid.org/0000-0000-9530-8833 (Fei Ding)

关键词: 周围神经损伤, 许旺细胞前体, 无细胞疗法, 条件培养基, 分泌组, 外囊泡, 微小RNA, 缺氧预处理, 氧糖剥夺, 轴突离断

Abstract: Our previous study found that rat bone marrow–derived neural crest cells (acting as Schwann cell progenitors) have the potential to promote long-distance nerve repair. Cell-based therapy can enhance peripheral nerve repair and regeneration through paracrine bioactive factors and intercellular communication. Nevertheless, the complex contributions of various types of soluble cytokines and extracellular vesicle cargos to the secretome remain unclear. To investigate the role of the secretome and extracellular vesicles in repairing damaged peripheral nerves, we collected conditioned culture medium from hypoxia-pretreated neural crest cells, and found that it significantly promoted the repair of sensory neurons damaged by oxygen-glucose deprivation. The mRNA expression of trophic factors was highly expressed in hypoxia-pretreated neural crest cells. We performed RNA sequencing and bioinformatics analysis and found that miR-21-5p was enriched in hypoxia-pretreated extracellular vesicles of neural crest cells. Subsequently, to further clarify the role of hypoxia-pretreated neural crest cell extracellular vesicles rich in miR-21-5p in axonal growth and regeneration of sensory neurons, we used a microfluidic axonal dissociation model of sensory neurons in vitro, and found that hypoxia-pretreated neural crest cell extracellular vesicles promoted axonal growth and regeneration of sensory neurons, which was greatly dependent on loaded miR-21-5p. Finally, we constructed a miR-21-5p-loaded neural conduit to repair the sciatic nerve defect in rats and found that the motor and sensory functions of injured rat hind limb, as well as muscle tissue morphology of the hind limbs, were obviously restored. These findings suggest that hypoxia-pretreated neural crest extracellular vesicles are natural nanoparticles rich in miRNA-21-5p. miRNA-21-5p is one of the main contributors to promoting nerve regeneration by the neural crest cell secretome. This helps to explain the mechanism of action of the secretome and extracellular vesicles of neural crest cells in repairing damaged peripheral nerves, and also promotes the application of miR-21-5p in tissue engineering regeneration medicine.

Key words: axotomy, cell-free therapy, conditioned medium, extracellular vesicles, hypoxic preconditioning, microRNA, oxygen-glucose deprivation, peripheral nerve injury, Schwann cell precursors