中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (2): 315-323.doi: 10.4103/1673-5374.265556

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

相对和绝对定量等量异位标签蛋白质组学分析脊神经Wallerian变性前后蛋白表达的变化

  

  • 出版日期:2020-02-15 发布日期:2020-05-25
  • 基金资助:

    国家重点研究发展计划（2016YFC11011601,2017YFA0104701），军事医学青年培养项目（15QNP091），解放军医学青年培训项目（16QNP144）


Different protein expression patterns in rat spinal nerves during Wallerian degeneration assessed using isobaric tags for relative and absolute quantitation proteomics profiling

Shuai Wei1, 2, 3, Xue-Zhen Liang2, 4, Qian Hu1, Wei-Shan Wang1, Wen-Jing Xu2, Xiao-Qing Cheng2, 3, Jiang Peng2, 3, Quan-Yi Guo2, Shu-Yun Liu2, Wen Jiang1, 2, 3, Xiao Ding1, 2, 3, Gong-Hai Han2, 3, 5, Ping Liu2, 3, 6, Chen-Hui Shi1, Yu Wang2, 3#br#   

  1. 1 The First Affiliated Hospital of Medical College, Shihezi University, Shihezi, Xinjiang Uygur Autonomous Region, China
    2 Institute of Orthopedics, Chinese PLA General Hospital, Beijing, China
    3 Co-Innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China
    4 The First Clinical Medical School, Shandong University of Traditional Chinese Medicine, Jinan, Shandong Province, China
    5 Kunming Medical University, Kunming, Yunnan Province, China
    6 Shanxi Medical University, Taiyuan, Shanxi Province, China
  • Online:2020-02-15 Published:2020-05-25
  • Contact: Yu Wang, MD,wangwangdian628@126.com; Chen-Hui Shi,1512905535@qq.com.
  • Supported by:
    This study was supported by National Key Research & Development Program of China, No. 2016YFC11011601, 2017YFA0104701, the Youth Cultivation Project of Military Medical Science, China, No. 15QNP091 (to YW), and People’s Liberation Army Youth Training Project for Medical Science of China, No. 16QNP144 (to YW).

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摘要：

周围神经中的感觉和运动神经纤维在损伤及再生过程的解剖学和功能变化不完全相同。为进一步揭示这些差异背后的生物学过程，(1)使用相对和绝对定量的等量异位标签离体的SD大鼠脊神经的感觉神经纤维以及运动神经纤维，以蛋白质组学技术分析其蛋白质谱的差异。结果显示在感觉神经中差异表达的蛋白有626种，其中368种上调，258种下调；而运动神经中差异表达的蛋白有637种，其中372种上调，265种下调；(2)通过生物信息学分析所有鉴定的蛋白，并以定量实时PCR对部分蛋白的mRNA表达进行验证，发现感觉神经中的差异表达蛋白主要与细胞-细胞粘附、碳水化合物代谢和细胞粘附等生物过程有关，而运动神经中的差异表达蛋白主要与糖酵解、细胞氧化还原稳态以及蛋白质折叠还原相关的生物过程有关；同时感觉神经和运动神经中的差异表达蛋白通常与细胞外泌体、髓鞘和黏着斑等细胞成分有关；(3)KEGG途径分析发现，差异表达蛋白主要与各种类型的代谢途径有关；(4)实验揭示了Wallerian变性过程中感觉神经和运动神经的差异表达蛋白，这将为未来感觉神经和运动神经Wallerian变性以及周围神经再生相关研究提供了一个蛋白质组学分析的数据库。实验于2016-09经中国人民解放军总医院伦理委员会批准（批准号2016-x9-07）。

orcid: 0000-0001-7811-5834 (Yu Wang)

关键词: 脊神经, 感觉神经, 运动神经, Wallerian变性, 基因本体论, KEGG途径, 相对和绝对定量的等量异位标签, 蛋白组学

Abstract: Sensory and motor nerve fibers of peripheral nerves have different anatomies and regeneration functions after injury. To gain a clear understanding of the biological processes behind these differences, we used a labeling technique termed isobaric tags for relative and absolute quantitation to investigate the protein profiles of spinal nerve tissues from Sprague-Dawley rats. In response to Wallerian degeneration, a total of 626 proteins were screened in sensory nerves, of which 368 were upregulated and 258 were downregulated. In addition, 637 proteins were screened in motor nerves, of which 372 were upregulated and 265 were downregulated. All identified proteins were analyzed using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis of bioinformatics, and the presence of several key proteins closely related to Wallerian degeneration were tested and verified using quantitative real-time polymerase chain reaction analyses. The differentially expressed proteins only identified in the sensory nerves were mainly relevant to various biological processes that included cell-cell adhesion, carbohydrate metabolic processes and cell adhesion, whereas differentially expressed proteins only identified in the motor nerves were mainly relevant to biological processes associated with the glycolytic process, cell redox homeostasis, and protein folding. In the aspect of the cellular component, the differentially expressed proteins in the sensory and motor nerves were commonly related to extracellular exosomes, the myelin sheath, and focal adhesion. According to the Kyoto Encyclopedia of Genes and Genomes, the differentially expressed proteins identified are primarily related to various types of metabolic pathways. In conclusion, the present study screened differentially expressed proteins to reveal more about the differences and similarities between sensory and motor nerves during Wallerian degeneration. The present findings could provide a reference point for a future investigation into the differences between sensory and motor nerves in Wallerian degeneration and the characteristics of peripheral nerve regeneration. The study was approved by the Ethics Committee of the Chinese PLA General Hospital, China (approval No. 2016-x9-07) in September 2016.

Key words: gene ontology, Kyoto Encyclopedia of Genes and Genomes, isobaric tags for relative and absolute quantitation, motor nerve, proteomics, sensory nerve, spinal nerve, Wallerian degeneration