title: Inhalation of activated protein C inhibits endotoxin-induced pulmonary inflammation in mice independent of neutrophil recruitment.
abstract: OBJECTIVE
  Intravenous administration of recombinant human activated protein C (rhAPC) is known to reduce lipopolysaccharide (LPS)-induced pulmonary inflammation by attenuating neutrophil chemotaxis towards the alveolar compartment. Ideally, one would administer rhAPC in pulmonary inflammation at the site of infection to minimize the risk of systemic bleeding complications. In this study, we therefore assessed the effect of inhaled rhAPC in a murine model of acute lung injury.
  
  
  METHODS
  Mice were exposed to LPS (0.5 mg kg(-1): intranasally) to induce acute lung injury. 30 minutes before and 3 hours after LPS exposure mice were subjected to vehicle or rhAPC inhalation (25 or 100 microg per mouse in each nebulization). In order to establish whether rhAPC inhalation affects neutrophil recruitment, neutrophil migration was determined in vitro using a trans-well migration assay.
  
  
  RESULTS
  rhAPC inhalation dose-dependently decreased LPS-induced coagulation and inflammation markers in bronchoalveolar lavage fluid (BALF), reduced protein leakage into the alveolar space and improved lung function. In contrast, rhAPC did not prevent LPS-induced neutrophil recruitment into the alveolar space. Neutrophil migration in vitro towards FCS or interleukin (IL)-8 was significantly inhibited by pretreatment with rhAPC (0.01-10 microg ml(-1)], whereas rhAPC (10 microg ml(-1)) added to the chemoattractant (modelling for topical rhAPC administration) did not affect neutrophil migration towards FCS or IL-8.
  
  
  CONCLUSIONS
  rhAPC inhalation significantly diminished LPS-induced pulmonary inflammation. The benefit of inhaled rhAPC appeared not to involve attenuation of neutrophil recruitment, in contrast to its effects after intravenous administration.
authors.affiliation: Center for Experimental and Molecular Medicine, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands. s.h.slofstra@amc.uva.nl
authors.forename: S H
authors.initials: SH
authors.lastname: Slofstra
authors.forename: A P
authors.initials: AP
authors.lastname: Groot
authors.forename: N A
authors.initials: NA
authors.lastname: Maris
authors.forename: P H
authors.initials: PH
authors.lastname: Reitsma
authors.forename: H Ten
authors.initials: HT
authors.lastname: Cate
authors.forename: C A
authors.initials: CA
authors.lastname: Spek
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects; D011014:Pneumonia / Q000517:prevention & control*; D011486:Protein C / Q000008:administration & dosage / Q000494:pharmacology*; D012129:Respiratory Function Tests
chemical_list: D008070:Lipopolysaccharides; D011486:Protein C