Department of Orthopaedics, General Hospital of Northern Theater Command, Shenyang, China.
Int J Exp Pathol. 2021 Aug;102(4-5):200-208. doi: 10.1111/iep.12400. Epub 2021 Oct 28.
MicroRNAs (miRNAs or miRs) serve essential roles in the pathogenic process of spinal cord injury (SCI). The present study investigated the role of miR-378-3p and autophagy-related 12 (ATG12) in SCI. RT-qPCR was used to detect the mRNA expression levels of miR-378-3p and ATG12. Cell viability and membrane integrity were evaluated using CCK-8 and LDH assays. For the analysis of the interaction between miR-378-3p and ATG12, a dual-luciferase reporter assay was conducted. The hindlimb function of rats was detected with the Basso, Beattie and Bresnahan score, and the motor deficit index score was used to evaluate nerve function. Using these approaches, it was identified that miR-378-3p expression was downregulated, while that of ATG12 was upregulated in SCI tissues and in cells exposed to hypoxia. Hypoxia repressed the expression of miR-378-3p via hypoxia-inducible factor 1-α. The overexpression of miR-378-3p exerted anti-apoptotic effects on nerve cells by directly repressing ATG12. The infusion of miR-378-3p improved hindlimb motor function and the neurological functions of rats with contusion SCI, which contributed to amelioration of functional deficits and the relief of contusion SCI. Therefore, it was concluded that upregulated expression of miR-378-3p in PC12 or N2A cells repressed the apoptosis of nerve cells, and the administration of miR-378-3p in model rats with contusion SCI improved neurological and motor functions.
微小 RNA(miRNA 或 miR)在脊髓损伤(SCI)的发病机制中发挥重要作用。本研究探讨了 miR-378-3p 和自噬相关 12(ATG12)在 SCI 中的作用。使用 RT-qPCR 检测 miR-378-3p 和 ATG12 的 mRNA 表达水平。使用 CCK-8 和 LDH 测定法评估细胞活力和膜完整性。为了分析 miR-378-3p 和 ATG12 之间的相互作用,进行了双荧光素酶报告基因测定。使用 Basso、Beattie 和 Bresnahan 评分检测大鼠后肢功能,并用运动缺陷指数评分评估神经功能。通过这些方法,发现 miR-378-3p 的表达在 SCI 组织和缺氧暴露的细胞中下调,而 ATG12 的表达上调。缺氧通过缺氧诱导因子 1-α 抑制 miR-378-3p 的表达。miR-378-3p 的过表达通过直接抑制 ATG12 对神经细胞发挥抗凋亡作用。miR-378-3p 的输注改善了挫伤 SCI 大鼠的后肢运动功能和神经功能,有助于改善功能缺陷并缓解挫伤 SCI。因此,结论是上调 PC12 或 N2A 细胞中 miR-378-3p 的表达可抑制神经细胞凋亡,并且在挫伤 SCI 模型大鼠中给予 miR-378-3p 可改善神经和运动功能。