title: Neutrophils from acute pancreatitis patients cause more severe in vitro endothelial damage compared with neutrophils from healthy donors and are differently regulated by endothelins.
abstract: OBJECTIVE
  There is evidence that endothelin (ET) 1 affect neutrophil functions and that patients with severe acute pancreatitis have increased plasma levels of ETs. Under appropriate conditions, neutrophils are able to injure the endothelium. In the present study, we compared healthy donors with acute pancreatitis patients for neutrophil degranulation and its ability to injure the endothelium and the contribution of ET-1 to this injury.
  
  
  METHODS
  Injury was evaluated by measuring the detachment of endothelial cells (ECV-304) growing in monolayer in coculture with human neutrophils for 4 hours. Neutrophil degranulation was assessed by myeloperoxidase (MPO) activity in coculture supernatants. In some experiments, neutrophils were pretreated with the antagonist of ET(A) receptor (BQ-123, 10(-6) M), which has high affinity for ET-1.
  
  
  RESULTS
  Neutrophils from both healthy donors and acute pancreatitis patients caused detachment of endothelial cells, and levels of MPO activity were increased in coculture supernatants. Neutrophils from acute pancreatitis patients caused significantly higher levels of detachment and MPO in the supernatants. Pretreatment of neutrophils with BQ-123 inhibited the detachment caused by neutrophils from healthy donors but not by neutrophils from acute pancreatitis patients.
  
  
  CONCLUSIONS
  These results show that neutrophils taken from healthy donors damage the endothelium by a mechanism dependent on ETs acting via ET(A) receptor, whereas neutrophils from acute pancreatitis patients cause more severe damage that is not dependent on ETs in the in vitro system used.
authors.affiliation: Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, Brazil.
authors.forename: Ellena C
authors.initials: EC
authors.lastname: Paulino
authors.forename: Lourenilson J
authors.initials: LJ
authors.lastname: de Souza
authors.forename: Nilza A T
authors.initials: NA
authors.lastname: Molan
authors.forename: Marcel C C
authors.initials: MC
authors.lastname: Machado
authors.forename: Sonia
authors.initials: S
authors.lastname: Jancar
mesh_terms: D000208:Acute Disease; D000328:Adult; D000959:Antihypertensive Agents / Q000494:pharmacology; D015550:Cell Degranulation / Q000276:immunology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000276:immunology / Q000473:pathology*; D065130:Endothelin A Receptor Antagonists; D019332:Endothelin-1 / Q000097:blood*; D006801:Humans; D066298:In Vitro Techniques; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D010195:Pancreatitis / Q000276:immunology* / Q000378:metabolism / Q000473:pathology*; D010456:Peptides, Cyclic / Q000494:pharmacology; D009195:Peroxidase / Q000378:metabolism; D044022:Receptor, Endothelin A / Q000378:metabolism*; D012720:Severity of Illness Index
chemical_list: D000959:Antihypertensive Agents; D065130:Endothelin A Receptor Antagonists; D019332:Endothelin-1; D010456:Peptides, Cyclic; D044022:Receptor, Endothelin A; D009195:Peroxidase; C072247:cyclo(Trp-Asp-Pro-Val-Leu)