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 (8): 1455-1461.doi: 10.4103/1673-5374.253531

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

坐骨神经损伤小鼠小泛素样修饰蛋白表达的多样性

  

  • 出版日期:2019-08-15 发布日期:2019-08-15
  • 基金资助:

    国家自然科学基金(316781246)；天津滨海新区卫生规划委员会科学技术基金(2011BHKY008，2012BWKZ002)；国家重点研究开发项目(2016YFC1101600)

Variation in expression of small ubiquitin-like modifiers in injured sciatic nerve of mice

Dian-Ying Zhang 1 , Kai Yu 2 , Zhong Yang 2 , Xiao-Zhi Liu 3 , Xiao-Fang Ma 3 , Yan-Xia Li 3   

  1. 1 Department of Orthopedics and Trauma, People’s Hospital, Peking University, Beijing, China
    2 Department of Orthopedics, Tianjin Fifth Central Hospital, Tianjin, China
    3 Central Laboratory, Tianjin Fifth Central Hospital, Tianjin, China
  • Online:2019-08-15 Published:2019-08-15
  • Contact: Dian-Ying Zhang, MSc, zdy8016@163.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (General Program), No. 316781246 (to DYZ); the Science and Technology Fund of Health Planning Commission of Binhai New Area of Tianjin of China, No. 2011BHKY008, 2012BWKZ002 (to KY); the National Key Research and Development Program of China, No. 2016YFC1101600 (to DYZ).

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摘要：

有研究发现小泛素样修饰蛋白(small ubiquitin-like modifier，SUMO)对神经生长、发育、分化过程中众多蛋白底物有共价可逆修饰作用，并参与神经轴突再生、信号传导、神经元迁移及髓鞘化的调控，但其在周围神经损伤修复过程中是否发挥作用以及作用机制，未见报道。为了解周围神经损伤修复过程中SUMO相关蛋白及SUMO化修饰过程有无潜在的神经保护作用，实验设计将C57/BL小鼠随机分成2组，假手术组仅暴露坐骨神经；模型组为坐骨神经离断、原位神经外膜缝合建立坐骨神经损伤修复模型。(1)采用Real-time PCR和western blot检测发现模型组不同取材时间点损伤神经组织(包括坐骨神经离断远近段分别5 mm组织)SUMO1，SUMO2，SUMO3和UBC9的mRNA和蛋白均有不同程度的表达，与假手术组相比，模型组不同取材时间点坐骨神经组织SUMO1、SUMO2/3蛋白在游离和结合状态均有表达，以结合状态尤为明显，而且模型组UBC9蛋白表达趋势与结合状态的SUMO1、SUMO2/3相一致；(2)免疫组织化学染色显示，模型组坐骨神经组织SUMO1及SUMO2/3免疫阳性反应均高于假手术组；(3)结果证实，坐骨神经损伤修复过程中，损伤神经组织可存在不同程度的SUMO1，SUMO2，SUMO3，UBC9的mRNA和蛋白表达，而SUMO相关蛋白的表达较为明显，从而揭示SUMOs可能在周围神经损伤修复中发挥重要作用。

orcid: 0000-0002-3204-4433 (Dian-Ying Zhang)

关键词: 周围神经损伤, 坐骨神经, SUMO1, SUMO2/3, UBC9, SUMO化修饰, 神经外膜缝合术, 神经再生

Abstract:

Small ubiquitin-like modifiers (SUMOs) have been shown to regulate axonal regeneration, signal transduction, neuronal migration, and myelination, by covalently and reversibly attaching to the protein substrates during neuronal cell growth, development, and differenti¬ation. It has not been reported whether SUMOs play a role in peripheral nerve injury and regeneration. To investigate any association between SUMOylation and potential neuroprotective effects during peripheral nerve injury and regeneration, C57/BL mice were ran¬domly divided into sham and experimental groups. The sciatic nerve was exposed only in the sham group. The experimental group underwent neurotomy and epineurial neurorrhaphy. Real-time quantitative polymerase chain reaction and western blot assay results revealed different mRNA and protein expression levels of SUMO1, SUMO2, SUMO3 and UBC9 in sciatic nerve tissue (containing both 5 mm of proximal and distal stumps at the injury site) at various time points after injury. Compared with the sham group, protein levels of SUMO1 and SUMO2/3 increased in both their covalent and free states after sciatic nerve injury in the experimental group, especially in the covalent state. UBC9 protein levels showed similar changes to those of SUMO1 and SUMO2/3 in the covalent states. Immunohisto¬chemical staining demonstrated that SUMO1 and SUMO2/3 immunopositivities were higher in the experimental group than in the sham group. Our results verified that during the repair of sciatic nerve injury, the mRNA and protein expression of SUMO1, SUMO2, SUMO3 and UBC9 in injured nerve tissues changed in varying patterns and there were clear changes in the expression of SUMO-related proteins. These findings reveal that SUMOs possibly play an important role in the repair of peripheral nerve injury. All animal protocols were ap¬proved by the Institutional Animal Care and Use Committee of Tianjin Fifth Central Hospital, China (approval No. TJWZXLL2018041) on November 8, 2018.

Key words: nerve regeneration, peripheral nerve injury, sciatic nerve, SUMO1, SUMO2/3, UBC9, SUMOylation, epineurial neurorrhaphy, neural regeneration