Genistein, a tyrosine kinase inhibitor, blocks the "second window of protection" 48 h after ischemic preconditioning in the rabbit.

We have previously reported a delayed or "second window of protection" against infarction 24-72 h after ischemic preconditioning in the rabbit. This phenomenon has also been associated with the protein kinase C signalling pathway. In the present study, we expanded our investigation to ascertain whether protein tyrosine kinase was in any way associated with this phenomenon in the rabbit heart. We found that 48 h after ischemic preconditioning with 4x5 min coronary occlusions the percentage of myocardium infarcting within the risk zone following a 30-min coronary occlusion and 120-min reperfusion (I/R) was reduced from 39. 6+/-3.3% to 18.0+/-3.7% (P<0.01). However, an i.v. bolus administration of genistein (5 mg/kg), a tyrosine kinase inhibitor, 5 min before ischemic preconditioning stimulus, abolished this protection (I/R=39.0+/-3.4%). Genistein per se had no significant effect on infarction 48 h later. Risk zone volume after coronary ligation was not significantly different between intervention groups. There were no differences in hemodynamic parameters between groups throughout the experimental period. We conclude that the second window of protection, 48 h after preconditioning, is mediated by tyrosine kinase activation in the rabbit heart.

Isolated dog hearts prepared in cold tyrode solution and reperfused with arterial blood are functionally and ultrastructurally normal.

Isolated dog whole heart preparations run by means of cross-circulation with support dogs were prepared in two different ways. In one way the donors' hearts were quickly removed, and cannulation of their major vessels, insertion of a latex balloon into the left ventricle, etc. were all done in cold Tyrode solution in the condition of heart-anoxia within 30 min and then reperfused with arterial blood of support dogs (Group A). In the other, cannulation of the major vessels of the donors' hearts were done in the state of "beating heart" (Group B). In these two kinds of preparations we compared basal cardio-coronary measurements (coronary blood flow, left ventricular pressure (LVP), its first derivative (LV dP/dt) and myocardial oxygen consumption (MVO2) and ultrastructure of the myocardium at various periods of the experiments. There were no significant differences in these cardio-coronary measurements and ultrastructural findings. Furthermore, Group A preparations kept stable levels of cardio-coronary measurements for 5 hr or more and responded normally to isoproterenol. Therefore, isolated, blood-perfused dog whole hearts prepared in cold Tyrode solution and reperfused by means of the cross-circulation method would be useful for physiological, pharmacological and biochemical studies.