中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (4): 812-818.doi: 10.4103/1673-5374.322467

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

CircLphn3具有保护创伤性脑损伤血脑屏障的作用

  

  • 出版日期:2022-04-15 发布日期:2021-10-18

CircLphn3 protects the blood-brain barrier in traumatic brain injury

Yu-Qi Cheng, Chen-Rui Wu, Meng-Ran Du, Qiang Zhou, Bi-Ying Wu, Jia-Yuan-Yuan Fu, Ehab Balawi, Wei-Lin Tan, Zheng-Bu Liao*   

  1. Department of Neurosurgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China
  • Online:2022-04-15 Published:2021-10-18
  • Contact: Zheng-Bu Liao, MD, liaozb123@cqmu.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81771355; and the Natural Science Foundation of Chongqing of China, No. CSTC2015jcyjA10096 (both to ZBL).

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摘要：

环状RNA是在中枢神经系统中大量表达的一大类新型非编码RNA分子。但是其对创伤性脑损伤的作用还知之甚少。（1）实验首先在创伤性脑损伤小鼠正常和损伤脑组织中筛选差异表达的环状RNA，发现circLphn3等环状RNA存在1倍及以上差异表达；（2）继而在创伤性脑损伤小鼠模型及血红素诱导的小鼠脑微血管内皮细胞（bEnd3）体外模型中，观察到circLphn3 mRNA和蛋白表达明显降低。而在bEnd3细胞中过表达circLphn3后，其紧密连接蛋白ZO1、ZO2和occludin表达上调，而miR-185-5p水平下调；（3）在转染miR-185-5p模拟物的bEnd3细胞中，ZO1表达水平下降；（4）以双萤光素酶报告系统分析，结果可见circLphn3可靶向miR-185-5p，而后者又可与ZO1结合。此外circLphn3过表达还能逆转血红素诱导的血脑屏障体外模型的高通透性，而miR-185-5p转染则加剧通透性损伤；（5）提示circLphn3可作为miR-185-5p的分子海绵，调节紧密连接蛋白的表达，从而改善血脑屏障的通透性。实验于2021年3月22日经重庆医科大学动物伦理委员会批准（批准号2021-177）。

https://orcid.org/0000-0003-4591-9689 (Zheng-Bu Liao); https://orcid.org/0000-0002-8281-2955 (Yu-Qi Cheng); 
https://orcid.org/0000-0002-4933-6918 (Chen-Rui Wu)

关键词: circRNA, 微小RNA, RNA测序, 生物信息学, ZO1, 紧密连接蛋白, 创伤性脑损伤, 血脑屏障

Abstract: Circular RNAs (circRNAs) are a new and large group of non-coding RNA molecules that are abundantly expressed in the central nervous system. However, very little is known about their roles in traumatic brain injury. In this study, we firstly screened differentially expressed circRNAs in normal and injured brain tissues of mice after traumatic brain injury. We found that the expression of circLphn3 was substantially decreased in mouse models of traumatic brain injury and in hemin-treated bEnd.3 (mouse brain cell line) cells. After overexpressing circLphn3 in bEnd.3 cells, the expression of the tight junction proteins, ZO-1, ZO-2, and occludin, was upregulated, and the expression of miR-185-5p was decreased. In bEnd.3 cells transfected with miR-185-5p mimics, the expression of ZO-1 was decreased. Dual-luciferase reporter assays showed that circLphn3 bound to miR-185-5p, and that miR-185-5p bound to ZO-1. Additionally, circLphn3 overexpression attenuated the hemin-induced high permeability of the in vitro bEnd.3 cell model of the blood-brain barrier, while miR-185-5p transfection increased the permeability. These findings suggest that circLphn3, as a molecular sponge of miR-185-5p, regulates tight junction proteins’ expression after traumatic brain injury, and it thereby improves the permeability of the blood-brain barrier. This study was approved by the Animal Care and Use Committee of Chongqing Medical University of China (approval No. 2021-177) on March 22, 2021. 

Key words: bioinformatics, blood-brain barrier, circRNA, miRNA, RNA-sequence, tight junction protein, traumatic brain injury, ZO1

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