Alamandine abrogates neutrophil degranulation in atherosclerotic mice

Eur J Clin Invest. 2017 Feb;47(2):117-128. doi: 10.1111/eci.12708. Epub 2017 Jan 4.

Abstract

Background: Neutrophil-mediated inflammation was recently identified as an active contributor to athero-progression. Therapeutic strategies inhibiting neutrophil degranulation or recruitment were hypothesized to positively impact on plaque vulnerability. In this study, we investigated whether treatment with the recently discovered agonist of the Mas-related G-coupled receptor type D (MrgD) alamandine would impact on neutrophil degranulation in vivo and in vitro.

Materials and methods: Fifteen-week-old ApoE-/- mice were fed with a Western-type diet for an additional 11 weeks. After the first 2 weeks of diet, mice were surgically implanted with a carotid 'cast' device that alters the blood shear stress and induces different carotid plaque phenotypes. During the last 4 weeks before euthanasia, mice were randomly assigned to subcutaneously receive vehicle (NaCl 0·15 M) or alamandine (24 μg/kg/h) by micropump. For in vitro experiments, neutrophils were obtained after thioglycollate intraperitoneal injection in ApoE-/- mice.

Results: Treatment with alamandine was well-tolerated, but failed to affect lipid, macrophage, neutrophil or collagen content within carotid and aortic root plaques. Also, treatment with alamandine did not affect Th-cell polarization in lymphoid organs. Conversely, alamandine administration was associated with a reduction in serum levels of neutrophil granule enzymes, such as MMP-9 and MPO as well as MMP-9 content within aortic root plaques. In vitro, preincubation with alamandine dose-dependently abrogated PMA-induced neutrophil degranulation of MMP-9 and MPO.

Conclusion: These results suggest that treatment with the MrgD agonist alamandine led to a reduced release of neutrophil granule products, potentially interfering with pro-atherosclerotic neutrophil activation.

Keywords: Atherosclerosis; Mas receptor; neutrophils.

MeSH terms

  • Animals
  • Aorta, Thoracic / drug effects
  • Atherosclerosis / drug therapy
  • Atherosclerosis / physiopathology*
  • Carotid Arteries / drug effects
  • Cell Degranulation / drug effects*
  • Disease Progression
  • In Vitro Techniques
  • Lipid Metabolism / drug effects
  • Matrix Metalloproteinase 9 / metabolism
  • Mice, Knockout
  • Neutrophils / drug effects*
  • Neutrophils / physiology
  • Oligopeptides / pharmacology*
  • Peroxidase / metabolism
  • Plaque, Atherosclerotic / drug therapy
  • Plaque, Atherosclerotic / physiopathology*
  • Random Allocation
  • Receptors, G-Protein-Coupled / agonists

Substances

  • Oligopeptides
  • Receptors, G-Protein-Coupled
  • alamandine
  • Peroxidase
  • Matrix Metalloproteinase 9

Associated data

  • GENBANK/NM_013488
  • GENBANK/NM_008176
  • GENBANK/NM_009140
  • GENBANK/NM_054039
  • GENBANK/NM_008091
  • GENBANK/NM_134250
  • GENBANK/NM_013556
  • GENBANK/NM_008337
  • GENBANK/NM_021283
  • GENBANK/NM_010548
  • GENBANK/NM_010552
  • GENBANK/NM_008611
  • GENBANK/NM_013599
  • GENBANK/NM_010824
  • GENBANK/NM_011281
  • GENBANK/NM_019471
  • GENBANK/NM_008605
  • GENBANK/NM_010809
  • GENBANK/NM_013693
  • GENBANK/NM_203490