中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (12): 2183-2191.doi: 10.4103/1673-5374.262602

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

胫神经和腓总神经损伤大鼠Wallerian变性过程中差异基因和蛋白的表达

  

  • 出版日期:2019-12-15 发布日期:2019-12-15
  • 基金资助:

    国家自然科学基金资助(81572146)；中国上海市卫生局优秀医疗人才计划(2017BR034)；上海教育发展基金会曙光计划和中国上海市教委项目(15SG34)

Differential gene and protein expression between rat tibial nerve and common peroneal nerve during Wallerian degeneration

Yao-Fa Lin 1 , Zheng Xie 1 , Jun Zhou 2 , Gang Yin 2 , Hao-Dong Lin 1, 2   

  1. 1 Department of Orthopedic Surgery, Changzheng Hospital, the Second Military Medical University, Shanghai, China
    2 Department of Orthopedic Surgery, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
  • Online:2019-12-15 Published:2019-12-15
  • Contact: Hao-Dong Lin, MD, PhD, linhaodong1978@smmu.ed
  • Supported by:

    This study was funded by the National Natural Science Foundation of China, No. 81572146 (to HDL); the Program of Outstanding Medical Talent of Shanghai Municipal Health Bureau, China, No. 2017BR034 (to HDL); the Shuguang Program of Shanghai Education Development Foundation and Shanghai Municipal Education Commission, China, No. 15SG34 (to HDL).

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摘要：

神经损伤后的Wallerian变性和再生是涉及多种细胞因子、基因和蛋白质的复杂过程。研究表明，不同周围神经损伤后再生速度不同，这是否是不同周围神经损伤后Wallerian变性过程中存在差异性基因和蛋白质导致的？具体的原因并不清楚。为此，实验设计了暴露大鼠右侧的胫神经和腓总神经，并在同一水平面完全切断后断端缝合，在术后第1，7，14和21天，分别切取大鼠手术侧的胫神经和腓总神经(缝合部位的远端神经组织，长1-2 cm)。采用高通量转录组测序(RNA-seq)和蛋白质组学技术研究大鼠模型中胫神经和腓总神经损伤后Wallerian变性过程中基因和蛋白表达的差异。(1)在术后第1，7，14和21天，在胫神经和腓总神经中，分别有1718，1374，1187和2195个差异表达基因表达，以及477，447，619和495个差异表达蛋白表达，在Wallerian变性过程中激活了47条通路；(2)实验筛选了在RNA-seq中有代表性的差异表达基因Hoxd4、Lpcat4、Tbx1进行RT-qPCR检测发现，基因表达变化与RNA-seq表达变化结果一致；(3)上述实验数据显示，在胫神经和腓总神经损伤后不同时间的Wallerian变性期间，两者远端神经组织中基因和蛋白质的表达有显著差异，说明不同类型周围神经损伤后Wallerian变性期间发生的生物过程是不同的。实验于2016年3月16日经中国第二军医大学动物伦理委员会批准(批准号：CZ20160218)。

orcid: 0000-0003-3461-3430(Hao-Dong Lin)

关键词: 周围神经, 周围神经损伤, Wallerian变性, 胫神经, 腓总神经, RNA测序, 蛋白质组学, 实时荧光定量PCR, SD大鼠, 神经再生

Abstract:

Wallerian degeneration and nerve regeneration after injury are complex processes involving many genes, proteins and cytokines. After different peripheral nerve injuries the regeneration rate can differ. Whether this is caused by differential expression of genes and proteins during Wallerian degeneration remains unclear. The right tibial nerve and the common peroneal nerve of the same rat were exposed and completely cut through and then sutured in the same horizontal plane. On days 1, 7, 14, and 21 after surgery, 1–2 cm of nerve tissue distal to the suture site was dissected out from the tibial and common peroneal nerves. The differences in gene and protein expression during Wallerian degeneration of the injured nerves were then studied by RNA sequencing and proteomic techniques. In the tibial and common peroneal nerves, there were 1718, 1374, 1187, and 2195 differentially expressed genes, and 477, 447, 619, and 495 differentially expressed proteins on days 1, 7, 14, and 21 after surgery, respectively. Forty-seven pathways were activated during Wallerian degeneration. Three genes showing significant differential expression by RNA sequencing (Hoxd4, Lpcat4 and Tbx1) were assayed by real-time quantitative polymerase chain reaction. RNA sequencing and real-time quantitative polymerase chain reaction results were consistent. Our findings showed that expression of genes and proteins in injured tibial and the common peroneal nerves were significantly different during Wallerian degeneration at different time points. This suggests that the biological processes during Wallerian degeneration are different in different peripheral nerves after injury. The procedure was approved by the Animal Experimental Ethics Committee of the Second Military Medical University, China (approval No. CZ20160218) on February 18, 2016.

Key words: nerve regeneration, peripheral nerves, peripheral nerve injuries, Wallerian degeneration, tibial nerve, common peroneal nerve, RNA sequencing, proteomic, real-time quantitative polymerase chain reaction, neural regeneration