Toll-like receptor 2/6 agonist macrophage-activating lipopeptide-2 promotes reendothelialization and inhibits neointima formation after vascular injury

Arterioscler Thromb Vasc Biol. 2013 Sep;33(9):2097-104. doi: 10.1161/ATVBAHA.113.301799. Epub 2013 Jul 18.

Abstract

Objective: Reendothelialization after vascular injury (ie, balloon angioplasty or stent implantation) is clinically extremely relevant to promote vascular healing. We here investigated the therapeutic potential of the toll-like receptor 2/6 agonist macrophage-activating lipopeptide (MALP)-2 on reendothelialization and neointima formation in a murine model of vascular injury.

Approach and results: The left common carotid artery was electrically injured, and reendothelialization was quantified by Evans blue staining after 3 days. A single injection of MALP-2 (1 or 10 µg, IV) after vascular injury accelerated reendothelialization (P<0.001). Proliferation of endothelial cells at the wound margins determined by 5-ethynyl-2'-deoxyuridine incorporation was significantly higher in MALP-2-treated animals (P<0.05). Furthermore, wire injury-induced neointima formation of the left common carotid artery was completely prevented by a single injection of MALP-2 (10 µg, IV). In vitro, MALP-2 induced proliferation (BrdU incorporation) and closure of an artificial wound of endothelial cells (P<0.05) but not of smooth muscle cells. Protein array and ELISA analysis of isolated primary endothelial cells and ex vivo stimulated carotid segments revealed that MALP-2 stimulated the release of multiple growth factors and cytokines predominantly from endothelial cells. MALP-2 induced a strong activation of the mitogen-activated protein kinase cascade in endothelial cells, which was attenuated in smooth muscle cells. Furthermore, MALP-2 significantly enhanced circulating monocytes and hematopoietic progenitor cells.

Conclusions: The toll-like receptor 2/6 agonist MALP-2 promotes reendothelialization and inhibits neointima formation after experimental vascular injury via enhanced proliferation and migration of endothelial cells. Thus, MALP-2 represents a novel therapeutic option to accelerate reendothelialization after vascular injury.

Keywords: cytokines; endothelial growth factors; neointima formation; toll-like receptors; vascular system injuries.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carotid Artery Injuries / drug therapy*
  • Carotid Artery Injuries / immunology
  • Carotid Artery Injuries / metabolism
  • Carotid Artery Injuries / pathology
  • Carotid Artery, Common / drug effects*
  • Carotid Artery, Common / immunology
  • Carotid Artery, Common / metabolism
  • Carotid Artery, Common / pathology
  • Cell Proliferation / drug effects*
  • Cells, Cultured
  • Cytokines / metabolism
  • Disease Models, Animal
  • Endothelial Cells / drug effects*
  • Endothelial Cells / immunology
  • Endothelial Cells / metabolism
  • Endothelial Cells / pathology
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Lipopeptides / pharmacology*
  • MAP Kinase Signaling System / drug effects
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mitogen-Activated Protein Kinases / metabolism
  • Myocytes, Smooth Muscle / drug effects
  • Myocytes, Smooth Muscle / immunology
  • Myocytes, Smooth Muscle / metabolism
  • Myocytes, Smooth Muscle / pathology
  • Neointima*
  • Platelet Aggregation / drug effects
  • Protein Array Analysis
  • Time Factors
  • Toll-Like Receptor 2 / agonists*
  • Toll-Like Receptor 2 / metabolism
  • Toll-Like Receptor 6 / agonists*
  • Toll-Like Receptor 6 / metabolism
  • Vascular System Injuries / drug therapy*
  • Vascular System Injuries / immunology
  • Vascular System Injuries / metabolism
  • Vascular System Injuries / pathology
  • Wound Healing / drug effects

Substances

  • Cytokines
  • Lipopeptides
  • Tlr2 protein, mouse
  • Tlr6 protein, mouse
  • Toll-Like Receptor 2
  • Toll-Like Receptor 6
  • macrophage stimulatory lipopeptide 2
  • Mitogen-Activated Protein Kinases