Pretreatment of brain dead rabbits with pinacidil before prolonged cold-storage with an extracellular solution alters aortic endothelial function

Abstract

Objective: Endothelial injury occurs during heart transplantation and contributes to the development of cardiac allograft vasculopathy. We have evaluated in a brain death model in the rabbit whether pre-treatment with the potassium channel opener (PCO) pinacidil before prolonged hypothermic storage with an extracellular solution would improve vascular endothelial recovery. Methods Rabbits were randomized into 4 experimental groups ( n = 8 per group). In the control group (CTRL), abdominal aortic rings were assessed immediately after 90 minutes of anesthesia. In the brain death group (BD), aortic rings were assessed immediately after 90 minutes of brain death. In the STH group, aortic rings taken from brain dead rabbits were stored for 24 hours at 4°C with the extracellular preservation solution of St. Thomas Hospital (STH) before assessment. In the STH + PCO group, the potassium channel opener pinacidil, 1 mg/kg, was administered intravenously to brain dead rabbits 10 minutes before explantation. Aortic rings were then stored for 24 hours at 4°C with the STH solution before evaluation. Brain death was induced by rapid inflation of a sub-durally placed balloon and validated by clinical and electroencephalographic data. Concentration-response curves to acetylcholine (ACH, 10 -9 to 10 -4 mol/liter) and nitroglycerin (NGL, 10 -9 to 10 -5 mol/liter) were constructed in phenylephrinepre-contracted rings. Results ACH evoked a similar concentration-dependent relaxation in the CTRL (E max: 95.8 ± 2.9%; EC 50: -6.86 ± 0.13 log M) and BD groups (E max: 90.8 ± 3.8%; EC 50: -6.75 ± 0.15 log M). The concentration-relaxation curve was shifted rightward in the STH group (E max: 76.7 ± 7.1%; EC 50: -6.75 ± 0.16 log M) in comparison with the CTRL and BD groups, but there were no significant differences in either E max or EC 50 values. After pinacidil pre-treatment, there was a further significant shift to the right of the concentration-relaxation curve to ACH (E max: 77.4 ± 5.0%; EC 50: -6.14 ± 0.19 log M, p < 0.05 vs CTRL, BD and STH). There were no significant differences between groups in the concentration-relaxation curves to NGL in endothelium-intact and endothelium-denuded vascular rings (either E max or EC 50). Conclusion Pre-treatment of brain dead rabbits with pinacidil before prolonged cold-storage with STH solution significantly impaired endothelium-dependent vasorelaxation in comparison to storage with STH solution. The role of PCO pre-treatment in the context of cardiac transplantation needs to be reconsidered.