MicroRNA-214-3p: A link between autophagy and endothelial cell dysfunction in atherosclerosis

Acta Physiol (Oxf). 2018 Mar;222(3). doi: 10.1111/apha.12973. Epub 2017 Oct 14.

Abstract

Aim: Endothelial cell injury assumes a fundamental part in the pathogenesis of atherosclerosis, and endothelial cell autophagy has protective effects on the development of atherosclerosis, although the underlying molecular regulation mechanism is indistinct. This study aimed to investigate whether microRNA-214-3p (miR-214-3p) is involved in the endothelial cell autophagy regulation of atherosclerosis.

Methods: We utilized ApoE-/- mice provided with a high-fat diet (HFD) as atherosclerosis model. We analysed the level of miR-214-3p and the levels of autophagy-related protein 5 (ATG5) and autophagy-related protein 12 (ATG12) in the purified CD31+ endothelial cells from mouse aorta. Bioinformatics analysis and a dual-luciferase reporter assay were performed to confirm the binding target of miR-214-3p. In vitro study, human umbilical vein endothelial cells (HUVECs) were transfected with miR-214-3p mimics/inhibitor and stimulated with 100 μg/mL oxidized low-density lipoprotein (ox-LDL) for 12 hours to initiate a stress-repairing autophagic process.

Results: In mouse models, we identified an inverse correlation between miR-214-3p, ATG5 and ATG12. We observed that in young HUVECs, ox-LDL-initiated autophagy was repressed by miR-214-3p overexpression, as evaluated by autophagic protein analysis, microtubule-associated protein 1 light chain 3B-II (LC3B-II) immunofluorescence assay and transmission electron microscopy (TEM). Also, miR-214-3p promoted ox-LDL accumulation in HUVECs and THP-1 monocyte adhesion. Conversely, in old HUVECs, suppression of miR-214-3p preserved the ability to initiate a protective autophagy reaction to the ox-LDL stimulation.

Conclusion: miR-214-3p regulates ox-LDL-initiated autophagy in HUVECs by directly targeting the 3'UTR of ATG5 and may have a suitable role in the pathogenesis of atherosclerosis.

Keywords: atherosclerosis; autophagy; human umbilical vein endothelial cells; miR-214-3p; oxidized low-density lipoprotein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Atherosclerosis / metabolism*
  • Autophagy / drug effects
  • Autophagy / physiology*
  • Diet, High-Fat
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Endothelial Cells / pathology*
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Lipoproteins, LDL / pharmacology
  • Mice
  • Mice, Knockout
  • MicroRNAs / metabolism*

Substances

  • Lipoproteins, LDL
  • MIRN214 microRNA, human
  • MicroRNAs
  • Mirn214 microRNA, mouse
  • oxidized low density lipoprotein