THE EFFECT OF OUABAIN (INHIBITOR OF AN ATP-DEPENDENT NA+ K+ PUMP) ON THE PANCREAS GRAFT DURING PRESERVATION BY THE 2-LAYER METHOD/

To clarify how synthesized ATP is utilized to maintain cellular integr ity during preservation by the two-layer method, we examined the effec t of ouabain, inhibitor of ATP dependent Na+/K+ pump, on ATP tissue le vels, graft weight, and vascular endothelial cells during 48 hr preser vation by the two-layer method and pancreatic tissue perfusion and gra ft survival after transplantation in a canine model, Ouabain treatment did not affect ATP production but prevented its utilization by the so dium pump, and actually significantly increased ATP levels and decreas ed the weight loss of the graft, In addition, ouabain caused a signifi cant increase of nuclear trypan-blue staining in vascular endothelial cells and a significant decrease of pancreatic tissue perfusion at rep erfusion. Consequently, the grafts did not survive, We conclude that t he two-layer method allows sufficient synthesis of ATP to drive the so dium pump and maintains membrane integrity of parenchymal cells and va scular endothelial cells, thus extending the period of preserved pancr eatic viability.