Decreased miR-146a expression in acute ischemic stroke directly targets the Fbxl10 mRNA and is involved in modulating apoptosis
Decreased miR-146a expression in acute ischemic stroke directly targets the Fbxl10 mRNA and is involved in modulating apoptosis
复制标题
急性缺血性中风中 miR-146a 表达降低直接靶向 Fbxl10 mRNA 并参与调节细胞凋亡
DOI：
10.1016/j.neuint.2017.01.011
复制
发表时间：
2017-07-01
影响因子：
4.2
通讯作者：
Liu, Jing-Li
中科院分区：
文献类型：
--
作者：
Li, Sheng-Hua;Chen, Lan;Liu, Jing-Li
背景：miR-146a 是某些病理生理过程中的强促凋亡因子，据报道与缺血性中风（IS）有关，但其作用尚不清楚。 Fbx110 是一种活性抗凋亡因子，也是 miR-146a 的预测靶标。我们假设 miR-146a 的失调通过靶向 Fbx110 导致缺血性损伤。方法：通过 miRNA 微阵列和 qRT-PCR 检测循环 miRNA。使用生物信息学预测 miR-146a 靶标，并通过双荧光素酶报告基因测定进行确认。我们使用氧糖剥夺和再灌注（OGD/R）的体外缺血模型来模拟脑缺血/再灌注（I/R）条件。通过 qRT-PCR 和蛋白质印迹验证 miR-146a、Fbx110 和 Bc1212 mRNA 以及 Fbx110 和 Bc1212 蛋白的表达。通过流式细胞术评估miR-146a对神经元细胞凋亡的影响。结果：在急性缺血性中风（AIS）中观察到miR-146a表达显着降低。双荧光素酶报告基因检测显示 Fbx110（而非 Bc1212）是 miR-146a 的靶标。转染 miR-146a 模拟物可促进 SK-N-SH 细胞凋亡并显着降低 Fbx110 的表达。相反，抑制miR-146a可减轻OGD/R诱导的神经细胞死亡，并显着上调Fbx110表达。结论：AIS中miR-146a表达显着下调，Fbx110被确定为miR-146a的靶标。此外，miR-146a 靶标 Fbx110 的上调可能可以保护神经元免于缺血性死亡。 (C) 2017 Elsevier Ltd. 保留所有权利。
Background: miR-146a, a strong pro-apoptotic factor in some pathophysiological processes, is reported to be involved in ischemic stroke (IS), though its role remains unclear. Fbx110 is an active anti-apoptotic factor and a predicted target of miR-146a. We hypothesized that dysregulation of miR-146a contributes to ischemic injury by targeting Fbx110.Methods: Circulating miRNAs were detected by miRNA microarray and qRT-PCR. miR-146a targets were predicted using bioinformatics and confirmed with a dual luciferase reporter assay. We used an in vitro ischemic model of oxygen-glucose deprivation and reperfusion (OGD/R) to mimic cerebral ischemia/reperfusion (I/R) conditions. Expression of miR-146a, Fbx110 and Bc1212 mRNAs, and Fbx110 and Bc1212 proteins was verified by qRT-PCR and Western blotting. The effects of miR-146a on neuronal cell apoptosis were evaluated by flow cytometry.Results: A significant reduction in miR-146a expression was observed in acute ischemic stroke (AIS). A dual-luciferase reporter assay showed that Fbx110, but not Bc1212, is a target of miR-146a. Transfection with miR-146a mimics promoted apoptosis in SK-N-SH cells and significantly reduced expression of Fbx110. Conversely, miR-146a inhibition attenuated OGD/R-induced neuronal cell death and significantly up-regulated Fbx110 expression.Conclusions: miR-146a expression was significantly down-regulated in AIS, and Fbx110 was identified as a target of miR-146a. Moreover, up-regulation of Fbx110, a miR-146a target, likely protects neurons from ischemic death. (C) 2017 Elsevier Ltd. All rights reserved.