MiR-590-5p-meidated LOX-1 upregulation promotes Angiotensin II-induced endothelial cell apoptosis
MiR-590-5p-meidated LOX-1 upregulation promotes Angiotensin II-induced endothelial cell apoptosis
复制标题
MiR-590-5p介导的LOX-1上调促进血管紧张素II诱导的内皮细胞凋亡
DOI：
10.1016/j.bbrc.2016.02.074
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发表时间：
2016-03-18
影响因子：
3.1
通讯作者：
Zhang, Zheng
中科院分区：
文献类型：
--
作者：
Luo, Ping;Zhang, Wei-Fang;Zhang, Zheng
背景：内皮细胞凋亡与高血压、动脉粥样硬化等心血管疾病的发生发展密切相关。microRNA调节内皮细胞功能，但其在内皮细胞凋亡中的作用仍有待充分阐明。方法：采用流式细胞术、Hoechst 33258染色和Western blotting方法检测血管紧张素II（Ang II）诱导的人脐静脉内皮细胞（HUVEC）凋亡。Western blotting和实时定量PCR检测LOX-1的表达。结果：Ang II诱导HUVEC凋亡时伴随miR-590- 5 p表达下调; miR-590 - 5 p模拟物可抑制HUVEC凋亡，减少ROS产生，表现为凋亡HUVEC比例减少和caspase-3活性降低。LOX-1的表达被Ang II增加，并且miR-590- 5 p模拟物在不存在或存在Ang II的情况下降低HUVEC中LOX-1的表达。用小干扰RNA或TS 92（LOX-1中和抗体）对LOX-1进行药理学或遗传阻断显著改善HUVEC凋亡，如通过减少凋亡HUVEC的数量、抑制caspase-3活化和抑制线粒体细胞色素C释放所证明的。结论：miR-590- 5 p下调可通过上调LOX-1的表达促进Ang II诱导的内皮细胞凋亡，从而增加ROS的产生。因此，miR-590- 5 p的恢复或LOX-1的阻断可以在治疗上用于减轻内皮细胞凋亡。(C)2016 Elsevier Inc. All rights reserved.
Background: Endothelial cell apoptosis contributes to cardiovascular diseases such as hypertension, atherosclerosis. MicroRNA regulates endothelial cell function but its role in endothelial cell apoptosis remains to be fully elucidated. This study aims to investigate the role of miR-590-5p in endothelial cell apoptosis and dissect the underlying mechanisms.Methods: Flow cytometric analysis, Hoechst 33258 staining and Western blotting were performed to evaluate human umbilical vein endothelial cell (HUVEC) apoptosis induced by Angiotensin (Ang) II. Western blotting and real-time quantitative PCR were conducted to assess the expression of LOX-1. DCFH-DA staining was carried out to measure the generation of reactive oxygen species (ROS).Results: Ang II-induced HUVEC apoptosis was accompanied by downregulation of miR-590-5p; administration of miR-590-5p mimics attenuated HUVEC apoptosis and decreased ROS generation, as indicated by reduced fraction of apoptotic HUVECs and decreased caspase-3 activity. LOX-1 expression was increased by Ang II, and miR-590-5p mimics reduced LOX-1 expression in HUVECs in the absence or presence of Ang II. Pharmacologic or genetic block of LOX-1 with small interference RNA or TS92 (LOX-1 neutralizing antibody) significantly ameliorated HUVEC apoptosis, as evidenced by reduced number of apoptotic HUVECs, inhibited caspase-3 activation and suppressed mitochondrial cytochrome C release. Moreover, LOX-1 siRNA or TS92 treatment dramatically reduced ROS production in HUVECs treated with Ang II.Conclusion: Our data demonstrated that miR-590-5p downregulation promoted Ang II-induced endothelial cell apoptosis by elevating LOX-1 expression and consequently increasing ROS generation. Thus, restoration of miR-590-5p or block of LOX-1 could be therapeutically exploited to alleviate endothelial cell apoptosis. (C) 2016 Elsevier Inc. All rights reserved.