中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (4): 889-894.doi: 10.4103/1673-5374.353504

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

大鼠脊髓损伤病理过程相关的循环外泌体lncRNA

  

  • 出版日期:2023-04-15 发布日期:2022-10-29

Circulating exosomal lncRNA contributes to the pathogenesis of spinal cord injury in rats

Jian-An Li1, #, Ming-Peng Shi2, #, Lin Cong3, Ming-Yu Gu2, Yi-Heng Chen2, Si-Yi Wang2, Zhen-Hua Li3, Chun-Fang Zan4, *, Wan-Fu Wei1, *   

  1. 1Department of Orthopedics, Tianjin Hospital, Tianjin, China;  2School of Clinical Medicine, Changchun University of Chinese Medicine, Changchun, Jilin Province, China;  3Department of Orthopedics, The Affiliated Hospital of Changchun University of Chinese Medicine, Changchun, Jilin Province, China;  4Division of Vascular Biology, Institute for Stroke and Dementia Research (ISD), LMU Klinikum, Ludwig-Maximilians-University (LMU), Munich, Germany
  • Online:2023-04-15 Published:2022-10-29
  • Contact: Chun-Fang Zan, PhD, chunfang.zan@med.uni-muenchen.de; Wan-Fu Wei, MD, weiwanfuortho@163.com.

摘要:

外泌体来源的长非编码RNAs(lncRNAs)可通过多种机制广泛参与到损伤脊髓的恢复和修复中。然而,到目前为止,还未有研究系统地评估与脊髓损伤(SCI)发展相关的差异表达lncRNAs。实验旨在筛选脊髓损伤大鼠中循环外泌体来源的几个关键lncRNAs,同时研究它们的潜在作用。为此,实验建立了大鼠脊髓半横断损伤模型。于脊髓损伤组和假手术对照组大鼠血液中提取循环外泌体。富集分析表明,脊髓损伤组和假手术对照组之间的lncRNAs和mRNAs表现出明显的不同表达模式。KEGG通路富集分析以及基因本体(GO)功能分析结果显示,上调和下调的lncRNAs可能涉及多种通路和生物过程、细胞定位和分子功能。定量反转录聚合酶链反应检测发现,包括ENSRN0T00000067908,XR_590093,XR_591455,XR_360081和XR_346933在内的5种lncRNAs表达增加,而包括XR_351404,XR_591426,XR_353833,XR_590076和XR_590719在内的其他5种lncRNAs下降。值得注意的是,lncRNA-miRNA-mRNA共表达网络显示,这10个lncRNA处于这个网络的中心,其中涉及198种mRNA和41种小干扰RNA。说明这些外泌体来源的lncRNAs可能参与了脊髓损伤的病理过程,其成为脊髓损伤的临床诊断和治疗的靶点。

https://orcid.org/0000-0002-4557-293X (Chun-Fang Zan); https://orcid.org/0000-0003-0729-9968 (Wan-Fu Wei)

Abstract: Exosome-derived long non-coding RNAs (lncRNAs) are extensively engaged in recovery and repair of the injured spinal cord, through different mechanisms. However, to date no study has systematically evaluated the differentially expressed lncRNAs involved in the development of spinal cord injury. Thus, the aim of this study was to identify key circulating exosome-derived lncRNAs in a rat model of spinal cord injury and investigate their potential actions. To this end, we established a rat model of spinal cord hemisection. Circulating exosomes were extracted from blood samples from spinal cord injury and control (sham) rats and further identified through Western blotting and electron microscopy. RNA was isolated from the exosomes and sequenced. The enrichment analysis demonstrated that there were distinctively different lncRNA and mRNA expression patterns between the two groups. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Ontology (GO) functional analysis were performed to determine the possible involvements of upregulated and downregulated lncRNAs in various pathways and different biological processes, as well as their cellular locations and molecular functions. Furthermore, quantitative reverse transcription-polymerase chain reaction showed that the expression of five lncRNAs––ENSRN0T00000067908, XR_590093, XR_591455, XR_360081, and XR_346933––was increased, whereas the expression of XR_351404, XR_591426, XR_353833, XR_590076, and XR_590719 was decreased. Of note, these 10 lncRNAs were at the center of the lncRNA-miRNA-mRNA coexpression network, which also included 198 mRNAs and 41 miRNAs. Taken together, our findings show that several circulating exosomal lncRNAs are differentially expressed after spinal cord injury, suggesting that they may be involved in spinal cord injury pathology and pathogenesis. These lncRNAs could potentially serve as targets for the clinical diagnosis and treatment of spinal cord injury.

Key words: exosome, inflammation, lncRNA, microenvironment, miRNA, mRNA, spinal cord injury, spinal cord repair