title: Myocardial apoptosis after cardioplegic arrest in the neonatal lamb.
abstract: OBJECTIVE
  Myocardial apoptosis is observed after various cardiac injuries and is also a normal part of fetal cardiac development and early postnatal maturation. Cardioplegic arrest and reperfusion result in ischemic injury and oxidative stress, known triggers of apoptosis. Because the neonatal heart is in a proapoptotic state, we hypothesize that apoptosis is triggered after cardioplegic arrest in neonatal myocardium.
  
  
  METHODS
  We started neonatal lambs (6-8 days old, n = 5) on cardiopulmonary bypass and administered cold crystalloid cardioplegia at 20-minute intervals. Total crossclamp time was 70 minutes, and bypass time was 90 minutes. After a six-hour recovery period, the hearts were excised and examined by using TdT-mediated dUTP nick-end labeling; radiolabeled DNA electrophoresis; fluorimetric caspase 3, 8, and 9 activity assay; mRNA microarray; and Western immunoblotting. Control lambs were anesthetized but did not undergo operation (n = 5) or were started on cardiopulmonary bypass for 90 minutes but not arrested (n = 5).
  
  
  RESULTS
  Lambs subjected to cardioplegia had 5-fold more TdT-mediated dUTP nick-end labeling-positive nuclei compared with that seen in unoperated control animals (P =.007) and bypass-only control animals (P =.008). DNA laddering was present in all postcardioplegia hearts but absent among control hearts. Bad and Bcl-X mRNA transcription increased significantly. Caspase 3, 8, and 9 activities were slightly greater than those seen in control animals, but the differences were not significant. No change was detected in Bcl-2, Bax, or Bcl-xL proteins.
  
  
  CONCLUSIONS
  In a clinically relevant model of neonatal cardioplegic arrest, increased apoptotic cell death is present 6 hours after reperfusion, and both proapoptotic and antiapoptotic responses are triggered. The clinical implications of apoptosis after cardioplegic arrest remain undetermined.
authors.affiliation: Department of Surgery, the University of Iowa College of Medicine, Iowa City, Iowa, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Hammel
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Caldarone
authors.forename: Timothy L
authors.initials: TL
authors.lastname: Van Natta
authors.forename: Li Xing
authors.initials: LX
authors.lastname: Wang
authors.forename: Karl F
authors.initials: KF
authors.lastname: Welke
authors.forename: Weigen
authors.initials: W
authors.lastname: Li
authors.forename: Scott
authors.initials: S
authors.lastname: Niles
authors.forename: Elisa
authors.initials: E
authors.lastname: Barner
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Scholz
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Behrendt
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Segar
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D020169:Caspases / Q000032:analysis; D006324:Heart Arrest, Induced / Q000009:adverse effects*; D020287:In Situ Nick-End Labeling; D009206:Myocardium / Q000473:pathology*; D040081:Protein Array Analysis; D012333:RNA, Messenger / Q000032:analysis; D012756:Sheep
chemical_list: D012333:RNA, Messenger; D020169:Caspases