中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (4): 764-769.doi: 10.4103/1673-5374.295335

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

长非编码RNA Airsci:增强大鼠脊髓损伤后的炎症反应

  

  • 出版日期:2021-04-15 发布日期:2020-12-22
  • 基金资助:

    国家自然科学基金(81801906);广东省科学基金(ZR2018PH024)

LncRNA Airsci increases the inflammatory response after spinal cord injury in rats through the nuclear factor kappa B signaling pathway

Tao Zhang1, 2, Kang Li2, Zi-Lu Zhang1, Kai Gao2, *, Chao-Liang Lv2, *   

  1. 1 School of Clinical Medicine, Jining Medical University, Jining, Shandong Province, China;  2 Department of Orthopedics, Jining No. 1 People’s Hospital, Jining, Shandong Province, China
  • Online:2021-04-15 Published:2020-12-22
  • Contact: Kai Gao, MD, gaohaikai88@126.com; Chao-Liang Lv, MD, lvchaolianggk@163.com.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China (No. 81801906; to KG) and the Natural Science Foundation of Shandong Province of China (No. ZR2018PH024; to KG).

摘要:

研究表明,长非编码RNA(LncRNA)在调节脊髓损伤急性期炎症反应的分子机制中起着重要作用。为探讨LncRNA Airsci通过核因子κB信号通路对脊髓损伤的作用机制,实验以LncRNA Airsci-siRNA 干预急性脊髓损伤。以Western Blot检测大鼠脊髓损伤后1-28d炎症因子核因子κB(p65)、核因子κB抑制因子、磷酸化核因子κB抑制因子的表达,发现伤后3d时核因子κB抑制因子核因子κB抑制因子表达达到峰值。通过RNA测序构建伤后3d的差异LncRNA表达谱,生物信息学技术分析差异表达的LncRNA,筛选出参与核因子κB信号通路并与急性炎症反应相关的LncRNA Airsci,最后通过实时荧光定量PCR进行验证。Western Blot和qRT-PCR检测脊髓损伤后第3天的损伤脊髓组织白细胞介素1β,白细胞介素6和肿瘤坏死因子α表达;用苏木精-伊红和尼氏染色法检测脊髓组织的病理改变;并通过BBB评分评估脊髓损伤大鼠的神经运动功能。结果表明,脊髓损伤后3d有151个LncRNA Airsci表达上调和186个LncRNA Airsci表达下调。LncRNA Airsci在涉及核因子κB信号通路的5个LncRNA中表达最明显。 LncRNA Airsci-siRNA 干预通过抑制核因子κB信号途径减轻炎症反应和脊髓组织损伤,并促进脊髓损伤大鼠运动功能的恢复。这些结果表明,抑制LncRNA Airsci可减轻炎症反应,促进脊髓损伤后的功能恢复,该作用机制涉及核因子κB信号通路。

https://orcid.org/0000-0002-0771-7755 (Kai Gao); 

https://orcid.org/0000-0002-4903-0345 (Chao-Liang Lv) 

Abstract: 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.