中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (11): 2257-2263.doi: 10.4103/1673-5374.310697

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

蛋白质组学鉴定脊髓源性星形胶质细胞重编程为神经元的关键蛋白

  

  • 出版日期:2021-11-15 发布日期:2021-04-13
  • 基金资助:

    福建省自然科学基金项目（2015J05153），福建省卫生厅科研人才培养项目（2018-ZQN-29），福建省科技创新联合基金（2018Y9002）

Identification of potential candidate proteins for reprogramming spinal cord-derived astrocytes into neurons: a proteomic analysis

Wen-Hao Chen, Yu-Xiang Lin, Ling Lin, Bao-Quan Zhang, Shu-Xia Xu, Wei Wang#br#   

  1. 1Department of Pediatric Surgery, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian Province, China; 2Department of Breast Surgery, Affiliated Union Hospital, Fujian Medical University, Fuzhou, Fujian Province, China; 3Institutes of Biomedical Sciences of Shanghai Medical School, Fudan University, Shanghai, China; 4Department of Neonatology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian Province, China; 5Department of Pathology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian Province, China; 6Department of Anatomy and Histoembryology, Fujian Medical University, Fuzhou, Fujian Province, China
  • Online:2021-11-15 Published:2021-04-13
  • Contact: Wen-Hao Chen, MD, 15606075620@163.com.
  • Supported by:
    The study was supported by the Natural Science Foundation of Fujian Province, China, No. 2015J05153; Research Talents Training Project of Fujian Provincial Health Department, China, No. 2018-ZQN-29; and Joint Funds for the Innovation of Science and Technology of Fujian Province, China, No. 2018Y9002 (all to WHC).

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摘要：

作者既往研究已证实，过表达Neurod1的脊髓源性星型胶质细胞可在在体条件下重编程为神经元，但其是否也可在体外条件下重编程为神经元，尚不可知，同时这一重编程的机制也不明确。（1）实验以刮伤法将来源于新生大鼠脊髓的星形胶质细胞制备成反应性星形胶质细胞，然后进行携带Neurod1的慢病毒转染。结果发现，Neurod1过表达可使体外培养的反应性星形胶质细胞重编程为神经元，其效率达13.4%；（2）基于蛋白质组学和生物信息学研究共鉴定出1952种蛋白，其中92种为差异表达蛋白。根据蛋白的生物功能和生物信息学倍数变化，确定出11种蛋白可能在这一重编程过程中可能起到关键作用；（3）经过Western blot验证，Neurod1过表达反应性星形胶质细胞中Csnk2a2和PNN表达显著提升，表明Neurod1可在体外直接将脊髓源性反应性星形胶质细胞重编程为神经元，且Neurod1-Csnk2a2-PNN通路可能参与这一过程。实验于2016年4月18日经福建医科大学动物伦理委员会批准（批准号2016-05）。

https://orcid.org/0000-0001-8988-7362 (Wen-Hao Chen)

关键词:

胶质细胞, 星形胶质细胞, 原代培养, 无标记蛋白质组学分析, 神经元, 重编程, 脊髓损伤, 机制

Abstract: Our previous study has confirmed that astrocytes overexpressing neurogenic differentiation factor 1 (NEUROD1) in the spinal cord can be reprogrammed into neurons under in vivo conditions. However, whether they can also be reprogrammed into neurons under in vitro conditions remains unclear, and the mechanisms of programmed conversion from astrocytes to neurons have not yet been clarified. In the present study, we prepared reactive astrocytes from newborn rat spinal cord astrocytes using the scratch method and infected them with lentivirus carrying NEUROD1. The results showed that NEUROD1 overexpression reprogrammed the cultured reactive astrocytes into neurons in vitro with an efficiency of 13.4%. Using proteomic and bioinformatic analyses, 1952 proteins were identified, of which 92 were differentially expressed. Among these proteins, 11 were identified as candidate proteins in the process of reprogramming based on their biological functions and fold-changes in the bioinformatic analysis. Furthermore, western blot assay revealed that casein kinase II subunit alpha (CSNK2A2) and pinin (PNN) expression in NEUROD1-overexpressing reactive astrocytes was significantly increased, suggesting that NEUROD1 can directly reprogram spinal cord-derived reactive astrocytes into neurons in vitro, and that the NEUROD1-CSNK2A2-PNN pathway is involved in this process. This study was approved by the Animal Ethics Committee of Fujian Medical University, China (approval No. 2016-05) on April 18, 2016. 

Key words: astrocytes, glial cells, label-free proteomic analysis, mechanism, neurons, primary culture, reprogramming, spinal cord injury

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