title: Lack of nuclear apoptosis in cardiomyocytes and increased endothelin-1 levels in a rat heart model of myocardial stunning.
abstract: OBJECTIVE
  Reperfusion injury may affect the cardiac NO and endothelin production. We investigated whether 20 min of total ischemia followed by 40 min of reperfusion can induce apoptosis in a Langendorff model of retrogradely perfused rat hearts (37 degrees C; paced at 300/'), and we attempted to correlate these findings with measured tissue NO and ET-1 levels.
  
  
  METHODS
  An apoptosis detection system was utilized which catalytically incorporates fluorescein-12-dUTP at the 3'-OH DNA ends using the principle of the TUNEL assay, with direct visualization of the labeled DNA. ET-1 was measured by radioimmunoassay and NO3/NO2 by ion pairing HPLC on C18 reverse phase columns.
  
  
  RESULTS
  None of the postischemic (n = 6) nor of the control perfused (90 min, n = 6) hearts showed signs of apoptosis, while those exposed to longer ischemia (40 min) and reperfusion (2 h) confirmed the presence of apoptotic cells. Myocardial ET-1 concentrations were 4.8 +/- 1.0 versus 8.3 +/- 2.5 pg/100 mg (control vs. ischemic hearts, respectively; mean +/- SD; p < 0.05). Myocardial NO contents showed no differences.
  
  
  CONCLUSIONS
  These data suggest that the time window of apoptosis with detectable DNA fragmentation exceeds 20 min of global total ischemia and 40 min of reperfusion, a model frequently used for inducing myocardial stunning. While NO was not increased in postischemic hearts, increased ET-1 levels indirectly argue for a role of ET-1 as inducer of apoptosis, but only at a later stage of reperfusion.
authors.affiliation: Div. of Cardiology, Zürich University Hospital.
authors.forename: M
authors.initials: M
authors.lastname: Klainguti
authors.forename: S
authors.initials: S
authors.lastname: Aigner
authors.forename: J
authors.initials: J
authors.lastname: Kilo
authors.forename: H M
authors.initials: HM
authors.lastname: Eppenberger
authors.forename: A
authors.initials: A
authors.lastname: Mandinova
authors.forename: U
authors.initials: U
authors.lastname: Aebi
authors.forename: M C
authors.initials: MC
authors.lastname: Schaub
authors.forename: S G
authors.initials: SG
authors.lastname: Shaw
authors.forename: T F
authors.initials: TF
authors.lastname: Lüscher
authors.forename: D
authors.initials: D
authors.lastname: Atar
mesh_terms: D000818:Animals; D017209:Apoptosis; D002467:Cell Nucleus / Q000473:pathology*; D004195:Disease Models, Animal; D019332:Endothelin-1 / Q000032:analysis / Q000096:biosynthesis*; D008854:Microscopy, Electron; D017202:Myocardial Ischemia / Q000378:metabolism / Q000473:pathology; D017682:Myocardial Stunning / Q000378:metabolism / Q000473:pathology*; D009206:Myocardium / Q000378:metabolism / Q000473:pathology*; D009569:Nitric Oxide / Q000032:analysis / Q000502:physiology; D051381:Rats; D011921:Rats, Inbred WKY; D013997:Time Factors; D016277:Ventricular Function, Left
chemical_list: D019332:Endothelin-1; D009569:Nitric Oxide