MicroRNA-214-3p: A link between autophagy and endothelial cell dysfunction in atherosclerosis
MicroRNA-214-3p: A link between autophagy and endothelial cell dysfunction in atherosclerosis
复制标题
MicroRNA-214-3p：动脉粥样硬化中自噬与内皮细胞功能障碍之间的联系
DOI：
10.1111/apha.12973
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发表时间：
2018-03-01
影响因子：
6.3
通讯作者：
Zhang, L.
中科院分区：
文献类型：
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作者：
Wang, J.;Wang, W. -N.;Zhang, L.
目的内皮细胞损伤在动脉粥样硬化的发病机制中发挥着重要作用，内皮细胞自噬对动脉粥样硬化的发展具有保护作用，但其分子调控机制尚不明确。本研究旨在探讨microRNA-214-3p（miR-214-3p）是否参与动脉粥样硬化的内皮细胞自噬调节。方法采用高脂饮食（HFD）的ApoE（-/-）小鼠作为动脉粥样硬化模型。我们分析了纯化的小鼠主动脉CD31+内皮细胞中miR-214-3p的水平以及自噬相关蛋白5（ATG5）和自噬相关蛋白12（ATG12）的水平。进行生物信息学分析和双荧光素酶报告基因测定以确认 miR-214-3p 的结合靶标。体外研究中，人脐静脉内皮细胞 (HUVEC) 转染 miR-214-3p 模拟物/抑制剂，并用 100 μg/mL 氧化低密度脂蛋白 (ox-LDL) 刺激 12 小时，以启动应激修复自噬过程。 结果在小鼠模型中，我们发现 miR-214-3p、ATG5 和 ATG12 之间存在负相关。我们观察到，在年轻的 HUVEC 中，ox-LDL 启动的自噬受到 miR-214-3p 过表达的抑制，通过自噬蛋白分析、微管相关蛋白 1 轻链 3B-II (LC3B-II) 免疫荧光测定和透射电子显微镜 (TEM) 进行评估。此外，miR-214-3p 促进 HUVEC 中 ox-LDL 的积累和 THP-1 单核细胞粘附。相反，在老的HUVEC中，抑制miR-214-3p保留了对ox-LDL刺激启动保护性自噬反应的能力。结论miR-214-3p通过直接靶向ATG5的3UTR来调节HUVEC中ox-LDL启动的自噬，并且可能在动脉粥样硬化的发病机制中发挥适当的作用。
AimEndothelial cell injury assumes a fundamental part in the pathogenesis of atherosclerosis, and endothelial cell autophagy has protective effects on the development of atherosclerosis, although the underlying molecular regulation mechanism is indistinct. This study aimed to investigate whether microRNA-214-3p (miR-214-3p) is involved in the endothelial cell autophagy regulation of atherosclerosis.MethodsWe utilized ApoE(-/-) mice provided with a high-fat diet (HFD) as atherosclerosis model. We analysed the level of miR-214-3p and the levels of autophagy-related protein 5 (ATG5) and autophagy-related protein 12 (ATG12) in the purified CD31(+) endothelial cells from mouse aorta. Bioinformatics analysis and a dual-luciferase reporter assay were performed to confirm the binding target of miR-214-3p. In vitro study, human umbilical vein endothelial cells (HUVECs) were transfected with miR-214-3p mimics/inhibitor and stimulated with 100 g/mL oxidized low-density lipoprotein (ox-LDL) for 12 hours to initiate a stress-repairing autophagic process.ResultsIn mouse models, we identified an inverse correlation between miR-214-3p, ATG5 and ATG12. We observed that in young HUVECs, ox-LDL-initiated autophagy was repressed by miR-214-3p overexpression, as evaluated by autophagic protein analysis, microtubule-associated protein 1 light chain 3B-II (LC3B-II) immunofluorescence assay and transmission electron microscopy (TEM). Also, miR-214-3p promoted ox-LDL accumulation in HUVECs and THP-1 monocyte adhesion. Conversely, in old HUVECs, suppression of miR-214-3p preserved the ability to initiate a protective autophagy reaction to the ox-LDL stimulation.ConclusionmiR-214-3p regulates ox-LDL-initiated autophagy in HUVECs by directly targeting the 3UTR of ATG5 and may have a suitable role in the pathogenesis of atherosclerosis.