Decreased miR‑92a‑3p expression potentially mediates the pro‑angiogenic effects of oxidative stress‑activated endothelial cell‑derived exosomes by targeting tissue factor.
Decreased miR‑92a‑3p expression potentially mediates the pro‑angiogenic effects of oxidative stress‑activated endothelial cell‑derived exosomes by targeting tissue factor.
复制标题
DOI：
10.3892/ijmm.2020.4713
复制
发表时间：
2020-11
影响因子：
5.4
通讯作者：
Chen H
中科院分区：
文献类型：
--
作者：
Li S;Yuan L;Su L;Lian Z;Liu C;Zhang F;Cui Y;Wu M;Chen H
血管生成是动脉粥样硬化易损斑块的基本病理特征。外泌体来源的microRNA（miRNAs或miRs）已被证明是血管生成的重要调节因子。然而，在氧化应激条件下由内皮细胞（EC）分泌的外泌体在血管生成中的作用仍不清楚。本研究旨在探讨氧化应激激活的内皮源性外泌体在血管生成中的作用及其机制。通过差速离心从H2 O2刺激的人脐静脉EC（HUVEC;命名）中分离外来体，并通过透射电子显微镜、纳米颗粒跟踪分析和蛋白质印迹分析进行表征。 增强HUVEC增殖、迁移和管形成，如分别通过EdU掺入测定、划痕伤口迁移测定和管形成测定所确定的。通过小RNA测序鉴定，miR-92 a-3 p是处理的HUVEC中主要下调的miRNA，并且如通过RT-qPCR所示，初级miR-92 a（pri-miR-92 a-1）的表达也降低。类似地，抑制miR-92 a-3 p在体外和体内促进血管生成。miR-92 a-3 p过表达阻断了对靶EC的促血管生成作用。组织因子（TF），一种参与血管生成的分子，在miR-92 a-3 p表达下调的HUVEC中增加，如mRNA测序所示。利用RNAhybrid程序预测TF是miR-92 a-3 p的靶点。miR-92 a-3 p的过表达或抑制在mRNA和蛋白质水平上修饰TF表达，如分别通过RT-qPCR和蛋白质印迹分析测量的。荧光素酶报告基因分析表明miR-92 a-3 p通过与TF的3′非翻译区结合抑制TF的表达。总之，本研究结果表明，氧化应激激活的内皮细胞释放的exosomes通过抑制受体内皮细胞中miR-92 a-3 p的表达来刺激血管生成，TF可能参与了miR-92 a-3 p对血管生成的调节作用。
Angiogenesis is an essential pathological feature of vulnerable atherosclerotic plaque. Exosome-derived microRNAs (miRNAs or miRs) have been proven to be important regulators of angiogenesis. However, the role of exosomes, which are secreted by endothelial cells (ECs) under conditions of oxidative stress, in angiogenesis remain unclear. The present study aimed to investigate the effects and mechanism of oxidative stress-activated endothelial-derived exosomes in angiogenesis. Exosomes were isolated from H2O2-stimulated human umbilical vein ECs (HUVECs; termed ) by differential centrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis and western blot analysis. enhanced HUVEC proliferation, migration and tube formation, as determined by EdU incorporation assay, scratch wound migration assay and tube formation assay, respectively. miR-92a-3p was identified as the predominantly downregulated miRNA in the -treated HUVECs by small RNA sequencing, and the expression of primary miR-92a (pri-miR-92a-1) was also decreased, as shown by RT-qPCR. Similarly, the inhibition of miR-92a-3p promoted angiogenesis in vitro and in vivo. miR-92a-3p overexpression blocked the pro-angiogenic effects of on target ECs. Tissue factor (TF), a molecule involved in angiogenesis, was increased in HUVECs in which miR-92a-3p expression was downregulated, as shown by mRNA sequencing. TF was also predicted as a target of miR-92a-3p by using the RNAhybrid program. The overexpression or suppression of miR-92a-3p modified TF expression at both the mRNA and protein level, as measured by RT-qPCR and western blot analysis, respectively. Luciferase reporter assays suggested that miR-92a-3p inhibited TF expression by binding to the 3′ untranslated region of TF. On the whole, the findings of the present study demonstrate that exosomes released from oxidative stress-activated ECs stimulate angiogenesis by inhibiting miR-92a-3p expression in recipient ECs, and TF may be involved in the regulatory effects of miR-92a-3p on angiogenesis.
登录
查看更多内容
影响因子：
5.3
作者：
Sun LL;Li WD;Lei FR;Li XQ
通讯作者：
Li XQ
MicroRNA-223抑制血管内皮细胞组织因子表达
影响因子：
5.3
作者：
Li, Sufang;Chen, Hong;Xu, Ning
通讯作者：
Xu, Ning
影响因子：
20.3
作者：
Deregibus, Maria Chiara;Cantaluppi, Vincenzo;Camussi, Giovanni
通讯作者：
Camussi, Giovanni
微泡介导的 microRNA-150 从单核细胞转移到内皮细胞可促进血管生成。
影响因子：
4.8
作者：
Li, Jing;Zhang, Yujing;Zhang, Chen-Yu
通讯作者：
Zhang, Chen-Yu
影响因子：
20.3
作者：
Umezu, Tomohiro;Tadokoro, Hiroko;Ohyashiki, Junko H.
通讯作者：
Ohyashiki, Junko H.