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 (3): 503-511.doi: 10.4103/1673-5374.266062

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

坐骨神经横断后远端细胞因子表达变化的蛋白质微阵列分析

  

  • 出版日期:2020-03-15 发布日期:2020-05-26
  • 基金资助:
    国家重点研究发展计划(2017YFA0104702），国家基础研究计划（973计划）（2014CB542201）

Protein microarray analysis of cytokine expression changes in distal stumps after sciatic nerve transection

Xiao-Qing Cheng1, Xue-Zhen Liang1, 2, Shuai Wei1, Xiao Ding1, Gong-Hai Han1, Ping Liu1, Xun Sun1, Qi Quan1, He Tang1, Qing Zhao1, Ai-Jia Shang3, 4, Jiang Peng1, 3   

  1. 1 Beijing Key Laboratory of Regenerative Medicine in Orthopedics, Institute of Orthopedics, Chinese PLA General Hospital, Beijing, China
    2 The First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan, Shandong Province, China
    3 Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China
    4 Department of Neurosurgery, Chinese PLA General Hospital, Beijing, China
  • Online:2020-03-15 Published:2020-05-26
  • Contact: Ai-Jia Shang, shangaj@126.com; Jiang Peng,pengjiang301@126.com.
  • Supported by:
    This study was supported by the National Key Research & Development Program of China, No. 2017YFA0104702 (to AJS), and the National Basic Research Program of China (973 Program), No. 2014CB542201 (to JP)

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摘要：

周围神经损伤后，大量趋化因子、细胞因子和其他营养因子以及细胞外基质形成利于周围神经再生的微环境，而这种微环境是再生成功的主要决定因素之一。因此，研究影响损伤后神经再生的关键分子和调节因子至关重要。目前。对坐骨神经损伤后不同时间点细胞因子表达的变化尚不明确。(1)实验通过横断坐骨神经，建立周围神经损伤模型，以蛋白质微阵列分析损伤后远端神经中不同细胞因子的表达，结果发现大量细胞因子在损伤后的不同时间点表达上调，但睫状神经营养因子等少量细胞因子出现下调。(2)以蛋白-蛋白相互作用网络筛选与差异表达的细胞因子相互作用的蛋白质，KEGG和基因本体分析结果表明，差异表达的细胞因子与趋化因子信号通路、JAK/STAT、PI3K/Akt以及notch信号通路有关。参与炎症和免疫反应以及细胞趋化性的细胞因子在神经损伤早期表达上调，参与神经细胞凋亡、细胞-细胞粘附和细胞增殖的细胞因子在损伤后28d时表达上调。(3)以Western blot验证肝细胞生长因子、胶质细胞源性神经营养因子和睫状神经营养因子在损伤后的表达及变化情况与蛋白质微阵列分析结果一致。(4) 结果能对全面了解细胞因子表达变化以及这些细胞因子与经典信号通路和生物学功能在Wallerian变性期间的变化提供帮助，同时有助于寻找周围神经损伤的潜在治疗方法。实验于2016年9月经中国人民解放军总医院机构动物护理和使用委员会批准（批准号2016-x9-07）。

orcid: 0000-0003-4662-9288 (Jiang Peng)

关键词: 神经远端, Wallerian变性, 蛋白质微阵列, 细胞因子, 蛋白-蛋白相互作用, KEGG通路, 基因本体, 周围神经损伤, 神经再生

Abstract: A large number of chemokines, cytokines, other trophic factors and the extracellular matrix molecules form a favorable microenvironment for peripheral nerve regeneration. This microenvironment is one of the major factors for regenerative success. Therefore, it is important to investigate the key molecules and regulators affecting nerve regeneration after peripheral nerve injury. However, the identities of specific cytokines at various time points after sciatic nerve injury have not been determined. The study was performed by transecting the sciatic nerve to establish a model of peripheral nerve injury and to analyze, by protein microarray, the expression of different cytokines in the distal nerve after injury. Results showed a large number of cytokines were up-regulated at different time points post injury and several cytokines, e.g., ciliary neurotrophic factor, were downregulated. The construction of a protein-protein interaction network was used to screen how the proteins interacted with differentially expressed cytokines. Kyoto Encyclopedia of Genes and Genomes pathway and Gene ontology analyses indicated that the differentially expressed cytokines were significantly associated with chemokine signaling pathways, Janus kinase/signal transducers and activators of transcription, phosphoinositide 3-kinase/protein kinase B, and notch signaling pathway. The cytokines involved in inflammation, immune response and cell chemotaxis were up-regulated initially and the cytokines involved in neuronal apoptotic processes, cell-cell adhesion, and cell proliferation were up-regulated at 28 days after injury. Western blot analysis showed that the expression and changes of hepatocyte growth factor, glial cell line-derived neurotrophic factor and ciliary neurotrophic factor were consistent with the results of protein microarray analysis. The results provide a comprehensive understanding of changes in cytokine expression and changes in these cytokines and classical signaling pathways and biological functions during Wallerian degeneration, as well as a basis for potential treatments of peripheral nerve injury. The study was approved by the Institutional Animal Care and Use Committee of the Chinese PLA General Hospital, China (approval number: 2016-x9-07) in September 2016.

Key words: cytokines, distal stump, gene ontology, Kyoto Encyclopedia of Genes and Genomes pathway, peripheral nerve injury, protein microarray, protein-protein interaction network, Wallerian degeneration