title: Leukocyte-depleted reperfusion after long cardioplegic arrest attenuates ischemia-reperfusion injury of the coronary endothelium and myocardium in rabbit hearts.
abstract: OBJECTIVE
  Cardiopulmonary bypass activates leukocytes, which should injure the coronary endothelium and myocardium during reperfusion especially after long cardioplegic arrest with long cardiopulmonary bypass time. The present study was designed to determine the protective efficacy of leukocyte-depleted reperfusion in blood-perfused parabiotic isolated rabbit hearts as a surgically relevant model with long cardioplegic arrest.
  
  
  METHODS
  Each isolated rabbit heart, with a latex balloon inserted in the left ventricle, was parabiotically blood-perfused using a modified Langendorff column. The left ventricular developed pressure (DP), rate of pressure development (dP/dT), and coronary flow with a left ventricular end-diastolic pressure of 10 mmHg were measured before ischemia and after 15, 30, 45, and 60 min reperfusion after 4 h cardioplegic arrest kept at 20 degrees C (control, n=10). Leukocyte-depleted reperfusion was done in the test group (n=10). The endothelium of the coronary artery was observed by scanning electron microscopy (SEM) with percent injured area of endothelial cells measured to evaluate the extent of endothelial ischemia-reperfusion injury.
  
  
  RESULTS
  The control hearts showed 53.3, 54.3, 48.4, and 39.0% recovery of DP compared to the pre-ischemia baseline data at 15, 30, 45, and 60 min after reperfusion began respectively. Leukocyte-depleted reperfusion enhanced the recovery of DP at 45 min (81.3%, P=0.0021) and 60 min (85.8%, P=0.0005) after reperfusion compared with that in the control group. The control hearts revealed 58.8%, 59.8%, 52.6%, and 43.4% recovery of dP/dT compared to the pre-ischemia baseline data at 15, 30, 45, and 60 min after reperfusion began, respectively. Leukocyte-depleted reperfusion also enhanced the recovery of dP/dT at 45 min (93.2%, P=0.0071) and 60 min (98.8%, P=0.0011) after reperfusion compared with that in the control group. There was also improvement of the recovery of coronary flow by leukocyte-depleted reperfusion (97.2%) compared with that in the control group (58.3%) after 60 min reperfusion (P=0.0121). Scanning electron microscopy showed that 69. 7% of coronary endothelial cells were morphologically injured in the control group. In contrast, leukocyte-depleted reperfusion prevented the extent of coronary endothelial damage with less injured area (0. 5%, P=0.0002).
  
  
  CONCLUSIONS
  Leukocyte-depleted reperfusion improved functional recovery with reduced coronary endothelial injury after long cardioplegic arrest.
authors.affiliation: Department of Thoracic and Cardiovascular Surgery, Saga Medical School, 5-1-1 Nabeshima, Saga City, 849-8501, Saga, Japan. okazaki@post.saga-med.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Okazaki
authors.forename: Z L
authors.initials: ZL
authors.lastname: Cao
authors.forename: S
authors.initials: S
authors.lastname: Ohtsubo
authors.forename: M
authors.initials: M
authors.lastname: Hamada
authors.forename: K
authors.initials: K
authors.lastname: Naito
authors.forename: K
authors.initials: K
authors.lastname: Rikitake
authors.forename: M
authors.initials: M
authors.lastname: Natsuaki
authors.forename: T
authors.initials: T
authors.lastname: Itoh
mesh_terms: D000818:Animals; D004730:Endothelium, Vascular / Q000648:ultrastructure*; D005112:Extracorporeal Circulation; D006324:Heart Arrest, Induced; D066298:In Vitro Techniques; D007962:Leukocytes; D008297:Male; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D009206:Myocardium / Q000648:ultrastructure*; D011817:Rabbits; D015424:Reperfusion / Q000379:methods*