Fu Xiaodong, Shen Yi, Wang Weili, Li Xiaomiao
Department of Orthopedics, School of Medicine, South Campus, Renji Hospital, Shanghai JiaoTong University, Shanghai, China.
Clin Exp Pharmacol Physiol. 2018 Jan;45(1):68-74. doi: 10.1111/1440-1681.12856. Epub 2017 Nov 6.
Spinal cord injury (SCI) is a major disability requiring more effective treatment than is currently available. MicroRNAs have been shown to effectively regulate gene expression at the translational level. The aim of the present study was to explore the potential role of miR-30-5p and possible mechanism in SCI. We found that miR-30-5p was notably down-regulated, while Neurod 1 expression was highly elevated in microglia from the mouse model of SCI. Additionally, overexpression of miR-30a-5p significantly suppressed inflammatory responses as reflected by a decrease in the secretion of the cytokines TNF-α, IL-1β and IL-10 triggered by SCI. Furthermore, introduction of miR-30a-5p strengthened the scavenging of oxygen free radicals accompanied by an increase in the expression of SEPN1, TXNL1 and GPX1. More importantly, our study explored that Neurod 1 was a direct and functional target of miR-30a-5p, which was validated by the dual luciferase reporter assay. qRT-PCR and western blot analysis further validated that miR-30a-5p negatively regulated the expression of Neurod 1. Mechanistically, overexpression of miR-30a-5p or silencing of the Neurod 1 gene prevented the MAPK/ERK signalling and inhibited inflammatory responses, meanwhile activated SEPN1, TXNL1 and GPX1. These findings indicate that miR-30a-5p ameliorates inflammatory responses and oxidative stress by targeting Neurod 1 through MAPK/ERK signalling.
脊髓损伤(SCI)是一种严重的残疾,需要比目前更有效的治疗方法。微小RNA已被证明能在翻译水平有效调节基因表达。本研究的目的是探讨miR-30-5p在脊髓损伤中的潜在作用及其可能机制。我们发现,在脊髓损伤小鼠模型的小胶质细胞中,miR-30-5p显著下调,而Neurod 1表达高度升高。此外,miR-30a-5p的过表达显著抑制了炎症反应,这表现为脊髓损伤引发的细胞因子TNF-α、IL-1β和IL-10分泌减少。此外,导入miR-30a-5p增强了氧自由基的清除,同时SEPN1、TXNL1和GPX1的表达增加。更重要的是,我们的研究发现Neurod 1是miR-30a-5p的直接功能性靶点,双荧光素酶报告基因检测验证了这一点。qRT-PCR和蛋白质印迹分析进一步验证了miR-30a-5p对Neurod 1表达的负调控作用。机制上,miR-30a-5p的过表达或Neurod 1基因的沉默可阻止MAPK/ERK信号传导并抑制炎症反应,同时激活SEPN1、TXNL1和GPX1。这些发现表明,miR-30a-5p通过MAPK/ERK信号传导靶向Neurod 1来改善炎症反应和氧化应激。