MicroRNA-223 inhibits tissue factor expression in vascular endothelial cells
MicroRNA-223 inhibits tissue factor expression in vascular endothelial cells
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MicroRNA-223抑制血管内皮细胞组织因子表达
DOI：
10.1016/j.atherosclerosis.2014.09.033
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发表时间：
2014-12-01
期刊：
影响因子：
5.3
通讯作者：
Xu, Ning
中科院分区：
文献类型：
--
作者：
Li, Sufang;Chen, Hong;Xu, Ning
目的：动脉粥样硬化是一种慢性炎症过程，血管内皮细胞（EC）由于炎症因子和生长因子等化学物质的作用而变得功能障碍。上述化学物质在活化的EC中诱导组织因子（TF）表达。 TF 在受损的动脉粥样硬化斑块上引发血栓形成，这在急性冠状动脉综合征 (ACS) 的发病过程中发挥着重要作用。越来越多的证据表明 microRNA 作为动脉粥样硬化疾病的表观遗传调节剂的重要作用。我们研究的目的是确定 microRNA-223 (miR-223) 是否靶向 EC 中的 TF。方法和结果：生物信息分析表明TF是miR-223的候选靶点。 Western blotting 分析显示，肿瘤坏死因子 α (TNF-α) 治疗 4 小时后增加了 C57BL/6J 小鼠主动脉和培养的 EC（EA.hy926 细胞和 HUVEC）的 TF 表达。在 TNF-α 处理的 EC 中，通过实时 PCR 测量，TF mRNA 也有所增加。实时 PCR 结果显示，经 TNF-α 处理的 C57BL/6J 小鼠主动脉和培养的 EC 中 miR-223 水平下调。用 miR-223 模拟物或 miR-223 抑制剂转染 EC 可以改变 mRNA 和蛋白质水平上的 TF 表达。荧光素酶测定证实，miR-223 通过与 TF 3'-非翻译区 (3'UTR) 序列结合来抑制 TF 表达。在有或没有 TNF-α 刺激的情况下，过表达 miR-223 可抑制 TF 促凝血活性。结论：MiR-223介导的TF表达抑制为凝血级联的调节提供了新的分子机制，并为动脉粥样硬化斑块破裂过程中血栓形成提供了线索。 (C) 2014 Elsevier Ireland Ltd. 保留所有权利。
Objective: Atherosclerosis is a chronic inflammatory process, in which vascular endothelial cells (ECs) become dysfunctional owing to the effects of chemical substances, such as inflammatory factor and growth factors. Tissue factor (TF) expression is induced by the above chemical substances in activated ECs. TF initiates thrombosis on disrupted atherosclerotic plaques which plays an essential role during the onset of acute coronary syndromes (ACS). Increasing evidences suggest the important role of microRNAs as epigenetic regulators of atherosclerotic disease. The aim of our study is to identify if microRNA-223 (miR-223) targets TF in ECs. Methods and results: Bioinformatic analysis showed that TF is a target candidate of miR-223. Western blotting analysis revealed that tumor necrosis factor alpha (TNF-alpha) increased TF expression in aorta of C57BL/6J mice and cultured ECs (EA.hy926 cells and HUVEC) after 4 h treatment. In TNF-alpha treated ECs, TF mRNA was also increased measured by real-time PCR. Real-time PCR results showed that miR-223 levels were downregulated in TNF-alpha-treated aorta of C57BL/6J mice and cultured ECs. Transfection of ECs with miR-223 mimic or miR-223 inhibitor modified TF expression both in mRNA and protein levels. Luciferase assays confirmed that miR-223 suppressed TF expression by binding to the sequence of TF 3'-untranslated regions (3'UTR). TF procoagulant activity was inhibited by overexpressing miR-223 with or without TNF-alpha stimulation. Conclusions: MiR-223-mediated suppression of TF expression provides a novel molecular mechanism for the regulation of coagulation cascade, and suggests a clue against thrombogenesis during the process of atherosclerotic plaque rupture. (C) 2014 Elsevier Ireland Ltd. All rights reserved.