中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (1): 179-185.doi: 10.4103/1673-5374.286979

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

Nogo-A加重了少突胶质细胞的氧化损害

  

  • 出版日期:2021-01-15 发布日期:2020-11-26
  • 基金资助:
    国家自然科学基金项目(81870996)

Nogo-A aggravates oxidative damage in oligodendrocytes

Yang-Yang Wang1, Na Han1, Dao-Jun Hong2, Jun Zhang1, *   

  1. 1 Department of Neurology, Peking University People’s Hospital, Beijing, China;   2 Department of Neurology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province, China
  • Online:2021-01-15 Published:2020-11-26
  • Contact: Jun Zhang, MD, who626@163.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 81870996 (to JZ).

摘要:

Nogo-A被认为是中枢神经系统中与髓鞘相关的最重要的轴突再生抑制剂之一,主要由少突胶质细胞表达。尽管既往研究已发现Nogo-A在神经突生长抑制、神经元稳态、神经前体细胞迁移、神经可塑性和神经退行性变中均能起到调节作用,但其在氧化损伤过程中的作用尚不明确。此次实验从SD乳鼠大脑皮质中提取少突胶质细胞,(1)以H2O2构建少突胶质细胞体外氧化损伤模型,发现少突胶质细胞中内源性Nogo-A的表达明显上调;(2)以重组病毒Ad-ZsGreen-Nogo-A转染少突胶质细胞后,Nogo-A表达受到抑制,发现这种细胞更容易受到急性氧化损伤,且细胞死亡增加。而以Ad-ZsGreen-shRNA-Nogo-A敲低少突胶质细胞中Nogo-A的表达后,外源H2O2引起的氧化应激明显减轻;(3)对少突胶质细胞加入Nogo-66-NgR/p75/LINGO-1-RhoA-ROCK通路抑制剂Nogo-66抗体、LINGO-1阻滞剂或Y27632抑制剂,不影响H2O2诱导的细胞死亡;(4) Ad-ZsGreen-shRNA-Nogo-A还可增加少突胶质细胞中磷酸化ERK1/2水平,并抑制Bcl-2的表达;(5)实验结果提示Nogo-A可加重少突胶质细胞的活性氧损害,且磷酸化ERKBcl-2可能参与了该过程。实验于2018-12-18经北京大学人民医院伦理委员会批准(批准号2018PHC081)。

https://orcid.org/0000-0002-5726-7190 (Jun Zhang)

关键词: Nogo-A, 少突胶质细胞, H2O2, 活性氧, 氧化损伤, Nogo-66-NgR/LINGO-1, 磷酸化ERK, Bcl-2

Abstract: Nogo-A is considered one of the most important inhibitors of myelin-associated axonal regeneration in the central nervous system. It is mainly expressed by oligodendrocytes. Although previous studies have found regulatory roles for Nogo-A in neurite outgrowth inhibition, neuronal homeostasis, precursor migration, plasticity, and neurodegeneration, its functions in the process of oxidative injury are largely uncharacterized. In this study, oligodendrocytes were extracted from the cerebral cortex of newborn Sprague-Dawley rats. We used hydrogen peroxide (H2O2) to induce an in vitro oligodendrocyte oxidative damage model and found that endogenously expressed Nogo-A is significantly upregulated in oligodendrocytes. After recombinant virus Ad-ZsGreen-rat Nogo-A infection of oligodendrocytes, Nogo-A expression was increased, and the infected oligodendrocytes were more susceptible to acute oxidative insults and exhibited a markedly elevated rate of cell death. Furthermore, knockdown of Nogo-A expression in oligodendrocytes by Ad-ZsGreen-shRNA-Nogo-A almost completely protected against oxidative stress induced by exogenous H2O2. Intervention with a Nogo-66 antibody, a LINGO1 blocker, or Y27632, an inhibitor in the Nogo-66-NgR/p75/LINGO-1-RhoA-ROCK pathway, did not affect the death of oligodendrocytes. Ad-ZsGreen-shRNA-Nogo-A also increased the levels of phosphorylated extracellular signal-regulated kinase 1/2 and inhibited BCL2 expression in oligodendrocytes. In conclusion, Nogo-A aggravated reactive oxygen species damage in oligodendrocytes, and phosphorylated extracellular signal-regulated kinase 1/2 and BCL2 might be involved in this process. This study was approved by the Ethics Committee of Peking University People’s Hospital, China (approval No. 2018PHC081) on December 18, 2018.

Key words: BCL2, H2O2, LINGO1, NgR, Nogo-A, oligodendrocytes, phosphorylated extracellular signal-regulated kinase 1/2, reactive oxygen species