miRNA-146a通过抑制IRAK1和TRAF6基因调控脊髓损伤炎症反应

发布时间：2018-04-05 08:31

本文选题：脊髓损伤　切入点：炎症反应　出处：《暨南大学》2017年博士论文

【摘要】：目的:近年来,越来越多的证据表明转录后的调控在机体功能发挥过程中发挥极其重要的作用。miRNA作为重要的表观遗传学内容受到越来越多的关注。目前与脊髓损伤(SCI)相关的miRNA研究大多数停留在生物信息学分析层面上,缺乏直接的生物学证据。我们拟在前期研究基础上,筛选与SCI炎症反应密切相关的miRNA,利用SCI动物模型以及临床样本探索miRNA在SCI炎症反应中的主要作用及其机制。材料与方法:利用SCI动物模型筛选目标miRNA,然后利用动物模型和临床SCI患者样本验证目标miRNA通过调控其靶基因参与SCI炎症反应并探索该过程的具体机理。同时检测分析SCI患者血液样本目标miRNA水平与SCI风险的相关性。结果:通过动物模型筛选发现miR-146a表达量在SCI后发生显著变化(P0.05)。发现miR-146a在SCI后3、7天表达量显著上调。炎症因子水平在SCI术后第1天显著上升,但在术后第3、7天,相对下降(P0.05)。形态学检测也发现在SCI术后损伤部位出现炎性细胞浸润。尾静脉注射miR-146a抑制剂,其靶基因IRAK1和TRAF6以及下游炎症因子的表达量显著升高(P0.05),炎症反应程度上升导致SCI恢复缓慢;反之,尾静脉注射miR-146a mimics则能够抑制其靶基因IRAK1和TRAF6以及下游炎症因子的表达量(P0.05),有利于SCI恢复;此外,实验还证实miR-146a能够显著抑制含有IRAK1和TRAF6基因3’UTR区荧光素酶载体的表达(P0.05)。SCI患者血液样本中miR-146a以及炎症因子表达规律与动物模型一致。结论:利用动物模型验证miR-146a在SCI后上调表达存在时间滞后性,首次提出miR-146a可能靶向下调IRAK1和TRAF6基因表达,同时又受到炎症因子的反馈调节,从而形成一个调控炎症反应的反馈调节机制,在SCI恢复过程发挥积极作用。miR-146a mimics可以降低炎症反应,一定程度上更好的促进SCI恢复,为SCI临床诊疗提供潜在治疗策略与治疗靶点。
[Abstract]:Objective: in recent years, more and more evidence that post transcriptional regulation plays an extremely important role in the process of.MiRNA as an important content of epigenetics has attracted more and more attention in the body function. At present with spinal cord injury (SCI) of miRNA related research most remain in the bioinformatics analysis level, the lack of direct biological evidence. We plan on the basis of former research, closely related to the miRNA screening and SCI inflammation, SCI animal models and clinical samples to explore the main role of miRNA in SCI in the inflammatory reaction and mechanism. Material and methods: the animal model of target miRNA by SCI screening, the specific mechanism and using animal models and clinical samples of patients with SCI verify that the target miRNA through the regulation of its target genes involved in the inflammatory reaction of SCI and explore the process. At the same time detection and analysis of blood samples of patients with SCI miRNA level target The correlation with the risk of SCI. Results: the animal models were found the expression of miR-146a was changed after SCI (P0.05). MiR-146a was found in the expression of 3,7 was up-regulated after SCI days. Significantly increased the level of inflammatory factors in the first day after SCI, but in 3,7 days after operation, the relative decline (P0.05). Morphological detection is also found in SCI after injury site of inflammatory cell infiltration. Intravenous injection of miR-146a inhibitor and its target gene IRAK1 and TRAF6 and the expression of inflammatory cytokines were significantly increased (P0.05), the degree of inflammatory reaction led to the rise in SCI recovery was slow; on the other hand, intravenous injection of miR-146a mimics can inhibit the target gene IRAK1 and TRAF6 and the expression of inflammatory cytokines content (P0.05), is conducive to the recovery of SCI; in addition, the experiment also confirmed the expression of miR-146a can inhibit the TRAF6 gene containing IRAK1 and 3 'UTR (P0.05).S luciferase vector Blood samples of patients with CI in miR-146a and the expression of inflammatory factors and rules consistent. Conclusion: the animal model using animal models to verify the miR-146a after SCI expression exists time lag, first proposed miR-146a may be targeted down-regulation of IRAK1 and TRAF6 gene expression, but also by the feedback regulation of inflammatory cytokines, thereby forming a regulation of inflammatory reaction a feedback regulation mechanism, play a positive role in SCI recovery process of.MiR-146a mimics can reduce the inflammatory response, to a certain extent, promote the recovery of SCI, and provide potential therapeutic target therapeutic strategy for clinical diagnosis and treatment of SCI.

【学位授予单位】：暨南大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R651.2

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