Department of Orthopedics, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
Department of Orthopedic, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, China.
J Orthop Res. 2022 Mar;40(3):661-673. doi: 10.1002/jor.25101. Epub 2021 Jun 6.
Long noncoding RNAs (lncRNAs) have been demonstrated to play critical regulatory roles in posttranscriptional and transcriptional regulation in eukaryotic cells. However, the characteristics of many lncRNAs, particularly their expression patterns in the lesion epicenter of spinal tissues following subacute spinal cord injury (SCI), remain unclear. In this study, we determined the expression profiles of lncRNAs in the lesion epicenter of spinal tissues after traumatic SCI and predicted latent regulatory networks. Standard Allen's drop surgery was conducted on mice, and hematoxylin and eosin staining was used to observe the damaged area. High-throughput sequencing was performed to identify the differential expression profiles of lncRNAs. Quantitative real-time polymerase chain reaction was conducted to evaluate the quality of the sequencing results. Bioinformatics analyses, including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, coexpression analysis, and protein-protein interaction analysis, were performed. Targeted binding of lncRNA-miRNA-mRNA was predicted by TargetScan and miRanda. A total of 230 differentially expressed lncRNAs were identified and preliminarily verified, and some potential regulatory networks were constructed. These findings improve our understanding of the mechanisms underlying subacute SCI; differentially expressed lncRNAs are closely involved in pathophysiological processes by regulating multiple pathways. Further studies are essential for revealing the exact mechanism underlying competing endogenous RNA pathways in vivo and in vitro.
长链非编码 RNA(lncRNA)已被证明在真核细胞中转录后和转录水平上发挥关键的调控作用。然而,许多 lncRNA 的特性,特别是它们在亚急性脊髓损伤(SCI)后脊髓组织损伤中心的表达模式,仍不清楚。在本研究中,我们确定了创伤性 SCI 后脊髓组织损伤中心 lncRNA 的表达谱,并预测了潜在的调控网络。对小鼠进行标准的 Allen 打击手术,并用苏木精和伊红染色观察损伤区域。进行高通量测序以鉴定 lncRNA 的差异表达谱。实时定量聚合酶链反应用于评估测序结果的质量。进行生物信息学分析,包括基因本体论和京都基因与基因组百科全书通路分析、共表达分析和蛋白质-蛋白质相互作用分析。通过 TargetScan 和 miRanda 预测 lncRNA-miRNA-mRNA 的靶向结合。共鉴定和初步验证了 230 个差异表达的 lncRNA,构建了一些潜在的调控网络。这些发现提高了我们对亚急性 SCI 机制的理解;差异表达的 lncRNA 通过调节多种途径密切参与病理生理过程。进一步的研究对于揭示体内和体外竞争内源性 RNA 途径的确切机制至关重要。