中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (2): 362-369.doi: 10.4103/1673-5374.317988

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

RNA干扰抑制EphB2表达可减弱胶质纤维瘢痕的形成并促进轴突生长 

  

  • 出版日期:2022-02-15 发布日期:2021-10-08

Downregulation of EphB2 by RNA interference attenuates glial/fibrotic scar formation and promotes axon growth

Jian Wu, Zhen-Yu Zhu, Zhi-Wei Fan, Ying Chen, Ri-Yun Yang, Yi Li*   

  1. Department of Histology and Embryology, Medical College, Nantong University, Nantong, Jiangsu Province, China
  • Online:2022-02-15 Published:2021-10-08
  • Contact: Yi Li, PhD, ly1203@ntu.edu.cn.
  • Supported by:
    This work was supported by the Priority Academic Program Development of Jiangsu Higher Education Institutes of China (PAPD); the Science and Technology Plan Project of Nantong of China, No. JC2020026 (to JW); the National Science Research of Jiangsu Higher Education Institutions of China, No. 19KJB310012 (to RYY)

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摘要：

胶质纤维瘢痕的快速形成是阻碍脊髓损伤后轴突生长的主要因素之一。成纤维细胞-星形胶质细胞接触依赖性相互作用中双向EphB2和ephrin-B2信号传导通路是胶质纤维瘢痕形成的触发因素。（1）此次实验将成纤维细胞和星形胶质细胞间隔培养在改良的载玻片系统并划痕损伤后制备了一种新的胶质纤维瘢痕体外模型，并在这种模型损伤之初使用RNA干扰技术抑制EphB2的表达后，观察胶质纤维瘢痕形成的变化以及VSC4.1运动神经元轴突生长情况；（2）结果可见干扰EphB2表达后，成纤维细胞和星形胶质细胞排列分散，不形成致密的胶质纤维瘢痕样结构，同时一些抑制轴突生长的细胞外基质成分neurocan，NG2和Ⅰ型胶原的表达明显减少，同时促进了微流控芯片中VSC4.1运动神经元轴突的生长；（3）结果说明RNAi抑制EphB2表达可抑制神经胶质纤维瘢痕的形成并促进轴突的生长。实验于2019年5月6日经江苏省实验动物伦理委员会批准（批准号2019-0506-002）。

https://orcid.org/0000-0002-7497-4823 (Yi Li)

关键词: EphB2, RNA干扰, 胶质纤维瘢痕, 脊髓损伤, 微流控芯片, 成纤维细胞, 星形胶质细胞, VSC4.1运动神经元

Abstract: The rapid formation of a glial/fibrotic scar is one of the main factors hampering axon growth after spinal cord injury. The bidirectional EphB2/ephrin-B2 signaling of the fibroblast-astrocyte contact-dependent interaction is a trigger for glial/fibrotic scar formation. In the present study, a new in vitro model was produced by coculture of fibroblasts and astrocytes wounded by scratching to mimic glial/fibrotic scar-like structures using an improved slide system. After treatment with RNAi to downregulate EphB2, changes in glial/fibrotic scar formation and the growth of VSC4.1 motoneuron axons were examined. Following RNAi treatment, fibroblasts and astrocytes dispersed without forming a glial/fibrotic scar-like structure. Furthermore, the expression levels of neurocan, NG2 and collagen I in the coculture were reduced, and the growth of VSC4.1 motoneuron axons was enhanced. These findings suggest that suppression of EphB2 expression by RNAi attenuates the formation of a glial/fibrotic scar and promotes axon growth. This study was approved by the Laboratory Animal Ethics Committee of Jiangsu Province, China (approval No. 2019-0506-002) on May 6, 2019.

Key words: astrocyte, EphB2, fibroblast, glial/fibrotic scar, microfluidic platform, RNAi, spinal cord injury, VSC4.1 motoneuron

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