中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (6): 1066-1070.doi: 10.4103/1673-5374.270314

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

氧糖剥夺大脑皮质神经元中前蛋白转化酶1/3可介导脑源性神经营养因子表达下调

  

  • 出版日期:2020-06-15 发布日期:2020-07-02
  • 基金资助:

    中国国家自然科学基金项目(81501053

Proprotein convertase 1/3-mediated down-regulation of brain-derived neurotrophic factor in cortical neurons induced by oxygen-glucose deprivation

Xiang-Yang Zhang1, Feng Liu1, Yan Chen2, Wei-Chun Guo1, Zhao-Hui Zhang2   

  1. 1 Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China
    2 Department of Neurology, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China
  • Online:2020-06-15 Published:2020-07-02
  • Contact: Yan Chen, PhD,chenyanzn@sina.com; Wei-Chun Guo, PhD,guoweichun@aliyun.com.
  • Supported by:
    This study was supported by the National Nature Science Foundation of China, No. 81501053 (to YC).

摘要:

脑源性神经营养因子对突触发生、神经元分化和突触传递和可塑性具有重要作用,但其成熟是一个复杂的过程。研究发现前蛋白转化酶1/3在受调节的分泌性蛋白前体的切割中起关键作用,其是否也参与缺血期间神经元中脑源性神经营养因子表达的变化,尚不得而知。为阐明前蛋白转化酶1/3在皮质神经元脑源性神经营养因子成熟中的作用,实验取胎鼠原代皮质神经元,经以无糖培养基5%CO2和95%N2潮湿密闭环境培养15min,37℃恢复3h后,ELISA和Western blot结果显示,氧糖氧糖剥夺后,神经元上清液以及胞内脑源性神经营养因子水平明显降低,且胞内前蛋白转化酶1/3水平下降。而氧糖氧糖剥夺后以pPC1/3质粒瞬时转染的皮质神经元中,当前蛋白转化酶1/3表达增加时,脑源性神经营养因子的表达也增加,且脑源性神经营养因子前体表达减少,上清液中脑源性神经营养因子浓度增加。结果表明在缺血过程中,前蛋白转化酶1/3对于皮质神经元中脑源性神经营养因子的转化至关重要。实验经武汉大学医学院动物伦理委员会批准。

orcid: 0000-0002-1424-1433 (Yan Chen)

关键词: 皮质神经元, 氧糖剥夺, 前蛋白转化酶1/3, 脑源性神经营养因子, 缺血, 前蛋白转化酶, 脑源性神经营养因子前体, 神经营养蛋白

Abstract: Brain-derived neurotrophic factor (BDNF) has robust effects on synaptogenesis, neuronal differentiation and synaptic transmission and plasticity. The maturation of BDNF is a complex process. Proprotein convertase 1/3 (PC1/3) has a key role in the cleavage of protein precursors that are directed to regulated secretory pathways; however, it is not clear whether PC1/3 mediates the change in BDNF levels caused by ischemia. To clarify the role of PC1/3 in BDNF maturation in ischemic cortical neurons, primary cortical neurons from fetal rats were cultured in a humidified environment of 95% N2 and 5% CO2 in a glucose-free Dulbecco’s modified Eagle’s medium at 37°C for 3 hours. Enzyme-linked immunosorbent assays and western blotting showed that after oxygen-glucose deprivation, the secreted and intracellular levels of BDNF were significantly reduced and the intracellular level of PC1/3 was decreased. Transient transfection of cortical neurons with a PC1/3 overexpression plasmid followed by oxygen-glucose deprivation resulted in increased PC1/3 levels and increased BDNF levels. When levels of the BDNF precursor protein were reduced, the concentration of BDNF in the culture medium was increased. These results indicate that PC1/3 cleavage of BDNF is critical for the conversion of pro-BDNF in rat cortical neurons during ischemia. The study was approved by the Animal Ethics Committee of Wuhan University School of Basic Medical Sciences.

Key words: cortical neuron, ischemia, neurotrophin, oxygen-glucose deprivation, precursor protein of brain-derived neurotrophic factor,
proprotein convertase,
proprotein convertase 1/3