title: Alamandine abrogates neutrophil degranulation in atherosclerotic mice.
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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Division of Cardiology, Department of Medical Specialties, Foundation for Medical Researches, University of Geneva, Geneva, Switzerland.
authors.forename: Analina R
authors.initials: AR
authors.lastname: Da Silva
authors.affiliation: Division of Cardiology, Department of Medical Specialties, Foundation for Medical Researches, University of Geneva, Geneva, Switzerland.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Lenglet
authors.affiliation: First Clinic of Internal Medicine, Department of Internal Medicine, University of Genoa School of Medicine, Genoa, Italy.
authors.forename: Federico
authors.identifier: http://orcid.org/0000-0003-2957-4078
authors.initials: F
authors.lastname: Carbone
authors.affiliation: Division of Cardiology, Department of Medical Specialties, Foundation for Medical Researches, University of Geneva, Geneva, Switzerland.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Burger
authors.affiliation: Division of Cardiology, Department of Medical Specialties, Foundation for Medical Researches, University of Geneva, Geneva, Switzerland.
authors.forename: Aline
authors.initials: A
authors.lastname: Roth
authors.affiliation: First Clinic of Internal Medicine, Department of Internal Medicine, University of Genoa School of Medicine, Genoa, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Liberale
authors.affiliation: First Clinic of Internal Medicine, Department of Internal Medicine, University of Genoa School of Medicine, Genoa, Italy.
authors.forename: Aldo
authors.initials: A
authors.lastname: Bonaventura
authors.affiliation: First Clinic of Internal Medicine, Department of Internal Medicine, University of Genoa School of Medicine, Genoa, Italy.|IRCCS AOU San Martino - IST, Genoa, Italy.
authors.forename: Franco
authors.initials: F
authors.lastname: Dallegri
authors.affiliation: Institute of Bioengineering, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Stergiopulos
authors.affiliation: Department of Physiology and Biophysics, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Robson A S
authors.initials: RA
authors.lastname: Santos
authors.affiliation: Division of Cardiology, Department of Medical Specialties, Foundation for Medical Researches, University of Geneva, Geneva, Switzerland.
authors.forename: François
authors.initials: F
authors.lastname: Mach
authors.affiliation: Institute of Bioengineering, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors.forename: Rodrigo A
authors.initials: RA
authors.lastname: Fraga-Silva
authors.affiliation: First Clinic of Internal Medicine, Department of Internal Medicine, University of Genoa School of Medicine, Genoa, Italy.|IRCCS AOU San Martino - IST, Genoa, Italy.|Centre of Excellence for Biomedical Research (CEBR), University of Genoa, Genoa, Italy.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Montecucco
mesh_terms: D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects; D050197:Atherosclerosis / Q000188:drug therapy / Q000503:physiopathology*; D002339:Carotid Arteries / Q000187:drug effects; D015550:Cell Degranulation / Q000187:drug effects*; D018450:Disease Progression; D066298:In Vitro Techniques; D050356:Lipid Metabolism / Q000187:drug effects; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D018345:Mice, Knockout; D009504:Neutrophils / Q000187:drug effects* / Q000502:physiology; D009842:Oligopeptides / Q000494:pharmacology*; D009195:Peroxidase / Q000378:metabolism; D058226:Plaque, Atherosclerotic / Q000188:drug therapy / Q000503:physiopathology*; D011897:Random Allocation; D043562:Receptors, G-Protein-Coupled / Q000819:agonists
chemical_list: D009842:Oligopeptides; D043562:Receptors, G-Protein-Coupled; C581752:alamandine; D009195:Peroxidase; D020780:Matrix Metalloproteinase 9
keywords: Atherosclerosis; Mas receptor; neutrophils