Zhang Tao, Li Kang, Zhang Zi-Lu, Gao Kai, Lv Chao-Liang
School of Clinical Medicine, Jining Medical University; Department of Orthopedics, Jining No. 1 People's Hospital, Jining, Shandong Province, China.
Department of Orthopedics, Jining No. 1 People's Hospital, Jining, Shandong Province, China.
Neural Regen Res. 2021 Apr;16(4):772-777. doi: 10.4103/1673-5374.295335.
Spinal cord injury (SCI) is a serious traumatic event to the central nervous system. Studies show that long non-coding RNAs (lncRNAs) play an important role in regulating the inflammatory response in the acute stage of SCI. Here, we investigated a new lncRNA related to spinal cord injury and acute inflammation. We analyzed the expression profile of lncRNAs after SCI, and explored the role of lncRNA Airsci (acute inflammatory response in SCI) on recovery following acute SCI. The rats were divided into the control group, SCI group, and SCI + lncRNA Airsci-siRNA group. The expression of inflammatory factors, including nuclear factor kappa B [NF-κB (p65)], NF-κB inhibitor IκBα and phosphorylated IκBα (p-IκBα), and the p-IκBα/IκBα ratio were examined 1-28 days after SCI in rats by western blot assay. The differential lncRNA expression profile after SCI was assessed by RNA sequencing. The differentially expressed lncRNAs were analyzed by bioinformatics technology. The differentially expressed lncRNA Airsci, which is involved in NF-κB signaling and associated with the acute inflammatory response, was verified by quantitative real-time PCR. Interleukin (IL-1β), IL-6 and tumor necrosis factor (TNF-α) at 3 days after SCI were measured by western blot assay and quantitative real-time PCR. The histopathology of the spinal cord was evaluated by hematoxylin-eosin and Nissl staining. Motor function was assessed with the Basso, Beattie and Bresnahan Locomotor Rating Scale. Numerous differentially expressed lncRNAs were detected after SCI, including 151 that were upregulated and 186 that were downregulated in the SCI 3 d group compared with the control group. LncRNA Airsci was the most significantly expressed among the five lncRNAs involved in the NF-κB signaling pathway. LncRNA Airsci-siRNA reduced the inflammatory response by inhibiting the NF-κB signaling pathway, alleviated spinal cord tissue injury, and promoted the recovery of motor function in SCI rats. These findings show that numerous lncRNAs are differentially expressed following SCI, and that inhibiting lncRNA Airsci reduces the inflammatory response through the NF-κB signaling pathway, thereby promoting functional recovery. All experimental procedures and protocols were approved by the approved by the Animal Ethics Committee of Jining Medical University (approval No. JNMC-2020-DW-RM-003) on January 18, 2020.
脊髓损伤（SCI）是一种对中枢神经系统造成严重创伤的事件。研究表明，长链非编码RNA（lncRNAs）在脊髓损伤急性期调节炎症反应中发挥重要作用。在此，我们研究了一种与脊髓损伤和急性炎症相关的新型lncRNA。我们分析了脊髓损伤后lncRNAs的表达谱，并探讨了lncRNA Airsci（脊髓损伤中的急性炎症反应）在急性脊髓损伤后恢复过程中的作用。将大鼠分为对照组、脊髓损伤组和脊髓损伤+lncRNA Airsci-siRNA组。通过蛋白质免疫印迹法检测大鼠脊髓损伤后1至28天炎症因子的表达，包括核因子κB [NF-κB (p65)]、NF-κB抑制剂IκBα和磷酸化IκBα（p-IκBα）以及p-IκBα/IκBα比值。通过RNA测序评估脊髓损伤后差异lncRNA表达谱。利用生物信息学技术分析差异表达的lncRNAs。通过定量实时PCR验证参与NF-κB信号传导并与急性炎症反应相关的差异表达lncRNA Airsci。通过蛋白质免疫印迹法和定量实时PCR检测脊髓损伤后3天的白细胞介素（IL-1β）、IL-6和肿瘤坏死因子（TNF-α）。通过苏木精-伊红染色和尼氏染色评估脊髓的组织病理学。用Basso、Beattie和Bresnahan运动评分量表评估运动功能。脊髓损伤后检测到大量差异表达的lncRNAs，与对照组相比，脊髓损伤3天组中有151个上调，186个下调。在参与NF-κB信号通路的5个lncRNAs中，lncRNA Airsci表达最为显著。LncRNA Airsci-siRNA通过抑制NF-κB信号通路减轻炎症反应，减轻脊髓组织损伤，并促进脊髓损伤大鼠运动功能的恢复。这些研究结果表明，脊髓损伤后大量lncRNAs差异表达，抑制lncRNA Airsci可通过NF-κB信号通路减轻炎症反应，从而促进功能恢复。所有实验程序和方案均于2020年1月18日获得济宁医学院动物伦理委员会批准（批准号：JNMC-2020-DW-RM-003）。
