title: Can the activation of plasminogen/plasmin system of the host by metabolic products of Dirofilaria immitis participate in heartworm disease endarteritis?
abstract: BACKGROUND
  Proliferative endarteritis is one of the key pathological mechanisms of cardiopulmonary dirofilariosis, a cosmopolitan parasitosis caused by Dirofilaria immitis affecting dogs and cats around the world. It has been shown that the excretory/secretory antigens from D. immitis adult worms (DiES) bind plasminogen (PLG) and activate fibrinolysis, which can lead to a survival mechanism for the parasite in its intravascular environment. However, overproduction of plasmin (final product of the route) has been related to pathological processes similar to those described in proliferative endarteritis. The aim of this study is to relate the appearance of this pathological condition with the activation of the PLG/plasmin system of the host by DiES.
  
  
  METHODS
  Cell proliferation through the crystal violet technique, cell migration by wound healing assay and degradation of the extracellular matrix by measuring collagen degradation and levels of matrix metalloproteinases were studied in an "in vitro" model using canine vascular endothelial and smooth muscle cells. These cells were treated with a mixture of DiES + PLG. Untreated cells, cells only stimulated with DiES or with PLG, or with a mixture of DiES + PLG + εACA (an inhibitor of the PLG-plasmin conversion) were employed as controls. In addition, the effect of DiES on the expression of the fibrinolytic activators tPA and uPA, the inhibitor PAI-1 and the PLG receptor Annexin A2 was analyzed in both types of cultures by western blot.
  
  
  RESULTS
  Plasmin generated by DiES + PLG binding produced a significant increase in the cell proliferation and migration of the endothelial and smooth muscle cells, as well as an increase in the destruction of the extracellular matrix based on a further degradation of Type I Collagen and an increased level of matrix metalloproteinase-2. DiES also induce an increase in the expression of tPA and uPA in endothelial cells in culture, as well as a decrease in the expression of PAI-1 in both types of cells.
  
  
  CONCLUSIONS
  Our study reports an interrelationship between plasmin caused by fibrinolysis activation by metabolic products of D. immitis and the appearance of pathological events similar to those described in the emergence of proliferative endarteritis in the cardiopulmonary dirofilariosis.
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Institute of Biomedical Research of Salamanca (IBSAL) and University of Salamanca, 37007, Salamanca, Spain. jglez@usal.es.
authors.forename: Javier
authors.initials: J
authors.lastname: González-Miguel
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Institute of Biomedical Research of Salamanca (IBSAL) and University of Salamanca, 37007, Salamanca, Spain. rmorgar@usal.es.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Morchón
authors.affiliation: Internal Medicine, Faculty of Veterinary Medicine, University of Las Palmas de Gran Canaria, 35413, Arucas, Las Palmas, Spain. elena.carreton@ulpgc.es.
authors.forename: Elena
authors.initials: E
authors.lastname: Carretón
authors.affiliation: Internal Medicine, Faculty of Veterinary Medicine, University of Las Palmas de Gran Canaria, 35413, Arucas, Las Palmas, Spain. alberto.montoya@ulpgc.es.
authors.forename: José Alberto
authors.initials: JA
authors.lastname: Montoya-Alonso
authors.affiliation: Laboratory of Parasitology, Faculty of Pharmacy, Institute of Biomedical Research of Salamanca (IBSAL) and University of Salamanca, 37007, Salamanca, Spain. fersimon@usal.es.
authors.forename: Fernando
authors.initials: F
authors.lastname: Simón
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003584:Cytological Techniques; D004183:Dirofilaria immitis / Q000378:metabolism / Q000502:physiology*; D004184:Dirofilariasis / Q000469:parasitology / Q000473:pathology*; D004195:Disease Models, Animal; D004285:Dogs; D004692:Endarteritis / Q000469:parasitology / Q000473:pathology*; D042783:Endothelial Cells / Q000469:parasitology / Q000473:pathology; D005341:Fibrinolysin / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000469:parasitology / Q000473:pathology; D010958:Plasminogen / Q000378:metabolism*; D010960:Plasminogen Activators / Q000378:metabolism*
chemical_list: D010958:Plasminogen; D010960:Plasminogen Activators; D005341:Fibrinolysin