中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (11): 2316-2323.doi: 10.4103/1673-5374.310700

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

大鼠面神经损伤修复分子机制的转录组学分析

  

  • 出版日期:2021-11-15 发布日期:2021-04-13

Transcriptome analysis of molecular mechanisms underlying facial nerve injury repair in rats

Qian-Qian Cao, Shuo Li, Yan Lu, Di Wu, Wei Feng, Yong Shi, Lu-Ping Zhang   

  1. 1Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China; 2Department of Otolaryngology, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China; 3Department of Otolaryngology, Head and Neck Surgery, Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China
  • Online:2021-11-15 Published:2021-04-13
  • Contact: Lu-Ping Zhang, PhD, zhanglp910@126.com; Yong Shi, PhD, yongshifudan@126.com.

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摘要：

既往多项微阵列分析已确定了面神经横断后面部运动神经元核中的转录组变化，但是损伤后面神经主干轴突内基因表达的改变尚不清楚。实验通过压迫成年大鼠右侧面神经颅外段主干建立了面神经挤压伤模型。（1）对损伤面部神经干行转录组测序以及差异基因表达分析和聚类分析，鉴定出39个存在特殊表达差异的交叉基因；（2）对这些交叉基因行GO和KEGG通路分析，发现这些基因主要参与白细胞粘附和吞噬作用，在调节神经修复中十分重要；（3）应用定量实时PCR验证关键基因的表达，确定了可能有助于面神经恢复的9个关键基因，Arhgap30，Akr1b8，C5ar1，Csf2ra，Dock2，Hcls1，Inpp5d，Sla和Spi1；（4）从新生大鼠坐骨神经中提取原代许旺细胞，以Akr1b8特异性siRNA敲除Akr1b8，可见许旺细胞中单核细胞趋化蛋白1和白细胞介素6的表达降低，而细胞增殖和迁移没有明显变化；（5）提示Akr1b8可能通过调节细胞因子表达而调控许旺细胞与巨噬细胞的相互作用促进面神经再生。研究首次揭示了损伤后面神经干的转录组变化，从而揭示了面神经损伤修复的潜在机制。研究于2018年经南通大学动物伦理委员会批准（批准号S20180923-007）。

https://orcid.org/0000-0003-1745-081X (Lu-Ping Zhang); https://orcid.org/0000-0003-4139-0116 (Yong Shi)

关键词:

面神经损伤, RNA序列, 转录组学分析, 许旺细胞, Akr1b8, 基因行为网络, 炎症反应, 细胞增殖

Abstract: Although the transcriptional alterations inside the facial nucleus after facial nerve injury have been well studied, the gene expression changes in the facial nerve trunk after injury are still unknown. In this study, we established an adult rat model of facial nerve crush injury by compressing the right lateral extracranial nerve trunk. Transcriptome sequencing, differential gene expression analysis, and cluster analysis of the injured facial nerve trunk were performed, and 39 intersecting genes with significant variance in expression were identified. Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the 39 intersecting genes revealed that these genes are mostly involved in leukocyte cell-cell adhesion and phagocytosis and have essential roles in regulating nerve repair. Quantitative real-time polymerase chain reaction assays were used to validate the expression of pivotal genes. Finally, nine pivotal genes that contribute to facial nerve recovery were identified, including Arhgap30, Akr1b8, C5ar1, Csf2ra, Dock2, Hcls1, Inpp5d, Sla, and Spi1. Primary Schwann cells were isolated from the sciatic nerve of neonatal rats. After knocking down Akr1b8 in Schwann cells with an Akr1b8-specific small interfering RNA plasmid, expression levels of monocyte chemoattractant protein-1 and interleukin-6 were decreased, while cell proliferation and migration were not obviously altered. These findings suggest that Akr1b8 likely regulates the interaction between Schwann cells and macrophages through regulation of cytokine expression to promote facial nerve regeneration. This study is the first to reveal a transcriptome change in the facial nerve trunk after facial nerve injury, thereby revealing the potential mechanism underlying repair of facial nerve injury. This study was approved by the Animal Ethics Committee of Nantong University, China in 2018 (approval No. S20180923-007). 

Key words: Akr1b8, cell proliferation, facial nerve injury, Gene-Act Networks, inflammatory response, RNA-Seq, Schwann cells, transcriptomics analysis

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