Research Paper Volume 11, Issue 9 pp 2762—2786

MiR-191 inhibit angiogenesis after acute ischemic stroke targeting VEZF1

Kang Du 1, 2, *, , Can Zhao 1, 2, *, , Li Wang 1, 2, , Yue Wang 1, 2, , Kang-Zhen Zhang 1, 2, , Xi-Yu Shen 1, 2, , Hui-Xian Sun 1, 2, , Wei Gao 1, 2, , Xiang Lu 1, 2, ,

  • 1 Department of Geriatrics, Sir Run Run Hospital, Nanjing Medical University, Nanjing, Jiangsu Province 211166, China
  • 2 Key Laboratory for Aging and Disease, Nanjing Medical University, Nanjing, Jiangsu Province 211166, China
* Equal contribution

received: February 15, 2019 ; accepted: April 29, 2019 ; published: May 7, 2019 ;

https://doi.org/10.18632/aging.101948
How to Cite

Copyright: Du et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Acute ischemic stroke (AIS) is a major public health problem in China. Impaired angiogenesis plays crucial roles in the development of ischemic cerebral injury. Recent studies have identified that microRNAs (miRNAs) are important regulators of angiogenesis, but little is known the exact effects of angiogenesis-associated miRNAs in AIS. In the present study, we detected the expression levels of angiogenesis-associated miRNAs in AIS patients, middle cerebral artery occlusion (MCAO) rats, and oxygen-glucose deprivation/reoxygenation (OGD/R) human umbilical vein endothelial cells (HUVECs). MiR-191 was increased in the plasma of AIS patients, OGD/R HUVECs, and the plasma and brain of MCAO rats. Over-expression of miR-191 promoted apoptosis, but reduced the proliferation, migration, tube-forming and spheroid sprouting activity in HUVECs OGD/R model. Mechanically, vascular endothelial zinc finger 1 (VEZF1) was identified as the direct target of miR-191, and could be regulated by miR-191 at post-translational level. In vivo studies applying miR-191 antagomir demonstrated that inhibition of miR-191 reduced infarction volume in MCAO rats. In conclusion, our data reveal a novel role of miR-191 in promoting ischemic brain injury through inhibiting angiogenesis via targeting VEZF1. Therefore, miR-191 may serve as a biomarker or a therapeutic target for AIS.

Abbreviations

AIS: acute ischemic stroke; BCA: Bradford protein assay; CCA: common carotid artery; CCK-8: cell counting kit-8; CITED2: Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 2; CO2: carbon dioxide; DMEM: Dulbecco's Modified Eagle Medium; ECA: external carotid artery; ECM: endothelial cell medium; EDN1: endothelin 1; EDTA: elhylene diamine tetraacetic acid; FBS: fetal bovine serum; FITC: fluorescein isothiocyanate; HRP: horseradish peroxidase; HUVEC: Human Umbilical Vein Endothelial Cell; ICA: internal carotid artery; MCAO: middle cerebral artery occlusion; miR-191: miR-191-5p; miR-191i: miR-191 inhibitor; miR-191m: miR-191 mimic; miRNA: microRNA; MMP: matrix metalloproteinase; MRI: magnetic resonance imaging; N2: nitrogen; NCi: negative control of inhibitor; NCm: negative control of mimic; O2: oxygen; OGD/R: oxygen-glucose deprivation/reoxygenatioin; PBS: phosphate buffer saline; PI: propidine iodide; qRT-PCR: quantitative real-time polymerase chain reaction; RNA: ribonucleic acid; STMN1: stathmin 1; TBP: TATA-box binding protein; TTC: 2,3,5-triphenyltetrazolium chloride; UTR: Un-translated region; VEGF-A: Vascular endothelial growth factor-A; VEZF1: vascular endothelial zinc finger 1.