title: Aprotinin impairs coronary endothelial function and down-regulates endothelial NOS in rat coronary microvascular endothelial cells.
abstract: OBJECTIVE
  The non-specific serine protease inhibitor aprotinin is currently used to reduce blood loss and the need for blood transfusion after cardiopulmonary bypass. We have recently reported that aprotinin impairs endothelium-dependent but not endothelium-independent relaxations in rat thoracic aortic rings due to its inhibitory effect on endothelial nitric oxide (NO) production. In light of these findings, the current study was designed to investigate the effects of aprotinin on coronary endothelial function in isolated rat hearts and on the expression of endothelial NO synthase (eNOS) in cultured rat coronary microvascular endothelial cells (CMEC).
  
  
  METHODS
  Hearts obtained from Sprague-Dawley rats were perfused on a constant flow Langendorff isolated heart system and coronary perfusion pressure and cardiac parameters were recorded. The coronary relaxant responses to bolus infusions of bradykinin (BK) and sodium nitroprusside (SNP) were recorded in the absence and presence of aprotinin. Total RNA and protein samples were extracted from CMEC incubated with aprotinin or the vehicle, 0.9% NaCl. RT-PCR-Southern blotting and Western analyses were carried out to assess eNOS mRNA and protein levels, respectively.
  
  
  RESULTS
  Aprotinin (125 and 250 kIU/ml) increased coronary perfusion pressure without changing the heart rate and cardiac contractility. Aprotinin inhibited BK-induced coronary vasodilatation at 250 kIU/ml, but not at 125 kIU/ml concentrations. The relaxant response to SNP did not change in response to either concentration of the drug. Incubation of CMEC with aprotinin down-regulated eNOS mRNA and protein at 250 kIU/ml, but not at 125 kIU/ml concentration.
  
  
  CONCLUSIONS
  These data suggest that aprotinin selectively inhibits NO synthesis at higher doses (> or = 250 kIU/ml) and therefore impairs endothelium-dependent coronary vascular tone. This effect of the drug may contribute to its 'blood-sparing' action, but may also account for the increase in the incidence of postoperative graft thrombosis observed in clinical practice during aprotinin therapy.
authors.affiliation: Department of Medicine, Institute of Clinical Science, Block B, Queen's University Belfast, Grosvenor Road, Belfast BT12 6BJ, Northern Ireland, UK.
authors.forename: Sibel
authors.initials: S
authors.lastname: Ulker
authors.forename: Pascal P
authors.initials: PP
authors.lastname: McKeown
authors.forename: Ulvi
authors.initials: U
authors.lastname: Bayraktutan
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000494:pharmacology*; D015139:Blotting, Southern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D003331:Coronary Vessels; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008833:Microcirculation; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D010477:Perfusion; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*
chemical_list: D012333:RNA, Messenger; D015842:Serine Proteinase Inhibitors; D009569:Nitric Oxide; D007611:Aprotinin; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat