Inhibition of heart transplant injury and graft coronary artery disease after prolonged organ ischemia by selective protein kinase C regulators.
OBJECTIVE
Transplanted hearts subjected to prolonged ischemia develop ischemia-reperfusion injury and graft coronary artery disease. To determine the effect of delta-protein kinase C and -protein kinase C on ischemia-reperfusion injury and the resulting graft coronary artery disease induced by prolonged ischemia, we used a delta-protein kinase C-selective inhibitor peptide and an -protein kinase C-selective activator peptide after 30 or 120 minutes of ischemia.


METHODS
Hearts of piebald viral glaxo (PVG) rats were heterotopically transplanted into allogeneic August Copenhagen Irish (ACI) rats. After cardioplegic arrest of the donor heart, -protein kinase C activator was injected antegrade into the coronary arteries. Hearts were procured and bathed in -protein kinase C activator, and before reperfusion, delta-protein kinase C inhibitor was injected into the recipient inferior vena cava. Controls were treated with saline. To analyze ischemia-reperfusion injury, grafts were procured at 4 hours after transplantation and analyzed for superoxide generation; myeloperoxidase activity; tumor necrosis factor alpha, interleukin 1beta, and monocyte/macrophage chemoattractant protein 1 production; and cardiomyocyte apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and caspase 2, 3, 8, and 9 activity. To analyze graft coronary artery disease, another set of animals underwent equal ischemic times and treatment strategies and then after 90 days were analyzed for graft coronary artery disease indexes.


RESULTS
All measures of ischemia-reperfusion injury and graft coronary artery disease after 120 minutes of ischemia in the saline-treated group were significantly increased relative to those observed after 30 minutes of ischemia. It is important to note that all ischemia-reperfusion injury parameters and graft coronary artery disease indexes decreased significantly in the protein kinase C regulator-treated group in comparison to saline-treated controls; additionally, these values were equivalent to those in saline-treated controls with 30 minutes of ischemia.


CONCLUSIONS
Combined treatment with -protein kinase C activator and delta-protein kinase C inhibitor reduces ischemia-reperfusion injury and decreases the resulting graft coronary artery disease induced by prolonged ischemia.
Tanaka
Masashi
M
Department of Cardiothoracic Surgery, Stanford University Medical Center, Falk Cardiovascular Research Center, Stanford, CA 94305, USA. masahi@omiya.jichi.ac.jp
Gunawan
Feny
F
Terry
Raya D
RD
Inagaki
Koichi
K
Caffarelli
Anthony D
AD
Hoyt
Grant
G
Tsao
Philip S
PS
Mochly-Rosen
Daria
D
Robbins
Robert C
RC
D000818:Animals; D017209:Apoptosis; D020169:Caspases / Q000032:analysis / Q000378:metabolism; D003327:Coronary Disease / Q000175:diagnosis / Q000209:etiology / Q000378:metabolism / Q000517:prevention & control*; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D004359:Drug Therapy, Combination; D004797:Enzyme-Linked Immunosorbent Assay; D006084:Graft Rejection; D016027:Heart Transplantation / Q000009:adverse effects*; D020287:In Situ Nick-End Labeling; D007249:Inflammation; D008297:Male; D015428:Myocardial Reperfusion Injury / Q000175:diagnosis / Q000209:etiology / Q000378:metabolism / Q000517:prevention & control*; D009206:Myocardium / Q000737:chemistry; D009842:Oligopeptides / Q000627:therapeutic use*; D009195:Peroxidase / Q000032:analysis / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology; D051744:Protein Kinase C-epsilon; D051381:Rats; D011919:Rats, Inbred Strains; D012720:Severity of Illness Index; D013481:Superoxides / Q000032:analysis / Q000378:metabolism; D013997:Time Factors; D016042:Transplantation, Heterotopic
C477271:His-Asp-Ala-Pro-Ile-Gly-Tyr-Asp; D009842:Oligopeptides; C477270:Ser-Phe-Asn-Ser-Tyr-Glu-Leu-Gly-Glu-Ser-Leu; D013481:Superoxides; D009195:Peroxidase; C497257:Prkce protein, rat; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D020169:Caspases