MiR-181b Antagonizes Atherosclerotic Plaque Vulnerability Through Modulating Macrophage Polarization by Directly Targeting Notch1

Mol Neurobiol. 2017 Oct;54(8):6329-6341. doi: 10.1007/s12035-016-0163-1. Epub 2016 Oct 8.

Abstract

Atherosclerotic plaque vulnerability is the major cause for acute stroke and could be regulated by macrophage polarization. MicroRNA-181b (miR-181b) was involved in macrophage differential. Here, we explore whether miR-181b could regulate atherosclerotic plaque vulnerability by modulating macrophage polarization and the underline mechanisms. In acute stroke patients with atherosclerotic plaque, we found that the serum level of miR-181b was decreased. Eight-week apolipoprotein E knockout (ApoE-/-) mice were randomly divided into three groups (N = 10): mice fed with normal saline (Ctrl), mice fed with high-fat diet, and tail vein injection with miRNA agomir negative control (AG-NC)/miR-181b agomir (181b-AG, a synthetic miR-181b agonist). We found that the serum level of miR-181b in AG-NC group was lower than that in Ctrl group. Moreover, 181b-AG could upregulate miR-181b expression, reduce artery burden and attenuate atherosclerotic plaque vulnerability by modulating macrophage polarization. In RAW264.7 cells treated with oxidized low-density lipoprotein (ox-LDL), we found miR-181b could reverse the function of ox-LDL on M1/M2 markers at both mRNA and protein levels. Furthermore, by employing luciferase reporter assay, we found that Notch1 was a direct target of miR-181b and could be regulated by miR-181b in vivo and in vitro. Finally, inhibition of Notch1 could abolish the function of downregulating miR-181b on increasing M2 phenotype macrophages. Our study demonstrates that administration of miR-181b could reduce atherosclerotic plaque vulnerability partially through modulating macrophage phenotype by directly targeting Notch1.

Keywords: Atherosclerotic plaque vulnerability; Ischemic stroke; Macrophage polarization; Notch1; miR-181b.

MeSH terms

  • Animals
  • Apolipoproteins E / genetics
  • Apolipoproteins E / metabolism
  • Cell Polarity / drug effects
  • Diet, High-Fat
  • Lipoproteins, LDL / pharmacology
  • Macrophages / drug effects*
  • Macrophages / metabolism
  • Mice
  • Mice, Knockout
  • MicroRNAs / agonists*
  • MicroRNAs / blood
  • Plaque, Atherosclerotic / metabolism*
  • Receptor, Notch1 / metabolism*

Substances

  • Apolipoproteins E
  • Lipoproteins, LDL
  • MicroRNAs
  • Notch1 protein, mouse
  • Receptor, Notch1
  • mirn181 microRNA, mouse
  • oxidized low density lipoprotein