MicroRNA-126 suppresses inflammation in endothelial cells under hyperglycemic condition by targeting HMGB1
MicroRNA-126 suppresses inflammation in endothelial cells under hyperglycemic condition by targeting HMGB1
复制标题
MicroRNA-126 通过靶向 HMGB1 抑制高血糖条件下内皮细胞的炎症
DOI：
10.1016/j.vph.2016.12.002
复制
发表时间：
2017-01-01
影响因子：
4
通讯作者：
Zhu, Hua-qing
中科院分区：
文献类型：
--
作者：
Tang, Song-tao;Wang, Feng;Zhu, Hua-qing
需要确定与炎症有关的microRNA-126(miR-126)靶点。在本研究中，我们旨在研究炎症相关基因高迁移率族蛋白1(HMGB1)是否为糖尿病血管内皮细胞miR-126的靶点。建立了经典的糖尿病动脉粥样硬化模型--糖尿病载脂蛋白E(-/-)小鼠模型。糖尿病apoE(-/-)小鼠主动脉miR-126表达降低，HMGB1表达升高，并伴有炎症反应。接下来，我们采用了几个体外实验来研究miRNA-126在高血糖和炎症条件下对内皮细胞HMGB1调节的作用。人脐静脉内皮细胞(HUVECs)中miRNA水平的调控是通过将miR-126模拟物和Anagomir基因导入细胞而实现的。在高血糖条件下，miR-126过表达可降低HUVECs HMGB1下游组分的表达，包括肿瘤坏死因子-α、ROS和NADPH氧化酶活性。然而，这种现象被miR-126 Anagomir完全逆转。经miR-126模拟载体转染后，HMGB1蛋白而不是HMGB1mRNA的表达下调，提示miR-126对HMGB1的调控作用处于翻译后水平。荧光素酶报告实验证实，HMGB1基因的3‘端非编码区是miR-126的直接靶点。Western印迹分析还表明，miR-126的过表达分别促进了p-eNOS、eNOS和p-ART的表达。综上所述，我们的研究结果提示miR-126可能通过调节HMGB1的表达来抑制高糖处理的内皮细胞的炎症和ROS的产生。我们的研究提供了糖尿病血管内皮细胞miRNA表达异常与炎症之间的新的致病联系。(C)2016 Elsevier Inc.保留所有权利。
MicroRNA-126(miR-126) targets involved in inflammation need to be identified. In this study, we aim to investigate whether high-mobility group box 1(HMGB1), an inflammation-related gene, is the target of miR-126 in diabetic vascular endothelium. The diabetic apoE(-/-) mice model, a classical diabetic atherosclerosis model, was established. The aorta of diabetic apoE(-/-) mice showed decrease of miR-126 and elevation of HMGB1 and inflammation. Next, we employed several in vitro experiments to address the role of miRNA-126 on the regulation of HMGB1 in endothelial cells under hyperglycemic and inflammatory conditions. Manipulation of miRNA levels in human umbilical vein endothelial cells (HUVECs) was achieved by transfecting cells with miR-126 mimic and antagomir. Overexpression of miR-126 could decrease the expression of downstream components of HMGB1 including TNF-alpha, ROS, and NADPH oxidase activity in HUVECs under hyperglycemic condition. Nevertheless, such phenomenon was completely reversed by miR-126 antagomir. The expression of HMGB1 protein rather than HMGB1 mRNA was down-regulated after transfection with miR-126 mimic, which indicated the modulation of HMGB1 mediated by miR-126 was at the posttranslational level. Luciferase reporter assay confirmed the 3'-UTR of HMGB1 gene was a direct target of miR-126. Western blot analysis also indicated that overexpression of miR-126 contributed to the elevation of p-eNOS, eNOS and p-ART expressions, respectively. In summary, our findings suggest that miR-126 may suppress inflammation and ROS production in endothelial cells treated by high glucose through modulating the expression of HMGB1. Our study provides a novel pathogenic link between dysregulated miRNA expression and inflammation in diabetic vascular endothelium. (C) 2016 Elsevier Inc. All rights reserved.