Endothelial enriched microRNAs regulate angiotensin II-induced endothelial inflammation and migration
Endothelial enriched microRNAs regulate angiotensin II-induced endothelial inflammation and migration
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DOI：
10.1136/heartjnl-2011-300867.133
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发表时间：
2011-04
期刊：
影响因子：
5.7
通讯作者：
Zhu Ni;W. Yuan;Sifen chen;Yongwen Qin
中科院分区：
文献类型：
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作者：
Zhu Ni;W. Yuan;Sifen chen;Yongwen Qin
动脉粥样硬化的初始阶段的特征在于白细胞的募集以激活内皮细胞（EC）。microRNA（miRNAs）是一类由19 ~ 25个核苷酸组成的非蛋白质编码RNA，通过翻译抑制或mRNA降解抑制靶基因的表达。miRNA和内皮功能之间的联系在很大程度上是未知的。北方杂交结果显示，miR-155和miR-221在人脐静脉内皮细胞（HUVECs）和血管平滑肌细胞（VSMCs）中高表达。生物信息学分析表明，Ets-1是炎症和管形成的关键内皮转录因子，是miR-155和miR-221/222簇的候选靶点。通过荧光素酶报告基因分析和Western blot证实了该作用。通过Western blot，我们也证实了血管紧张素II 1型受体（AT 1 R）是miR-155在HUVECs中的作用靶点。定量PCR结果显示Ets-1及其下游基因VCAM 1、MCP 1和FLT 1在血管紧张素II刺激的HUVECs中表达上调，而miR-155和miR-221/222的过表达可部分逆转Ets-1及其下游基因VCAM 1、MCP 1和FLT 1的表达。此外，细胞粘附实验显示过表达miR-155和miR-221/222可有效降低Jurkat T细胞与Ang II刺激的HUVECs的粘附。此外，miR-155还可以通过靶向AT 1 R，抑制血管紧张素II诱导的HUVECs迁移。综上所述，高表达miR-155的HUVEC可能共同靶向AT 1 R和Ets-1，而miR-221/222靶向Ets-1，其间接调节EC的几种炎症分子的表达，从而减弱Jurkat T细胞与活化的HUVEC的粘附，减少HUVEC的迁移。这些发现为动脉粥样硬化的治疗提供了可能的靶点。
The initial stage of atherosclerosis is characterised by recruitment of leukocytes to activate endothelial cells (ECs). MicroRNAs (miRNAs) are a class of 19 to 25 nucleotides, non-protein-coding RNAs that repress target gene expression by translational inhibition or mRNA degradation. The link between miRNA and endothelial functions is largely unknown. Northern blot showed that miR-155 and miR-221 were highly expressed in human umbilical vein endothelial cells (HUVECs) and vascular smooth muscle cells (VSMCs). Bioinformatics analysis proposed Ets-1, a key endothelial transcription factor for inflammation and tube formation, as a candidate target for miR-155 and miR-221/222 cluster. The effect was demonstrated by luciferase reporter assay and Western blot. By using Western blot, we also confirmed that angiotensin II type 1 receptor (AT1R) is a target of miR-155 in HUVECs. Quantitative PCR showed that Ets-1 and its downstream genes, including VCAM1, MCP1 and FLT1, were up-regulated in angiotensin II-stimulated HUVECs, and this effect was partially reversed by over-expression of miR-155 and miR-221/222. In addition, cell adhesion assay revealed over-expression of miR-155 and miR-221/222 effectively decreased the adhesion of Jurkat T cells to Ang II-stimulated HUVECs. Besides, by targeting AT1R, miR-155 can also decrease the HUVECs migration in response to Ang II. In summary, HUVECs highly expressed miR-155 may co-target AT1R and Ets-1 while miR-221/222 targets Ets-1, which indirectly regulate the expression of several inflammatory molecules of ECs, and therefore attenuate the adhesion of Jurkat T cells to activated HUVECs and reduce HUVECs migration. These findings present possible therapeutic targets in atherosclerosis.