Ductal Injection of JNK Inhibitors Before Pancreas Preservation Prevents Islet Apoptosis and Improves Islet Graft Function

Abstract

Human islet isolation to obtain high-quality islets is still challenging. This study investigates how c-Jun NH2- terminal kinase (JNK ) is activated during human and porcine islet isolation. We also investigated whether ductal injection of preservation solution with JNK inhibitors improves islet isolation results by preventing apoptosis of islet cells. A low molecular weight inhibitor (SP600125) and a cell-permeable peptide inhibitor, the latter introduced by protein transduction technology, were used in porcine and human studies, respectively. JNK activity progressively increased during the isolation procedure. The addition of 10 microM JNK inhibitors into the ductal preservation solution prevented JNK activation during the isolation procedure and prevented islet apoptosis immediately after isolation. We incubated islets (2000 islet equivalents) for 24-48 hr and then transplanted them below the kidney capsule of streptozotocin-induced diabetic mice. The blood glucose levels reached normoglycemia in more than 80% of the JNK inhibitor-positive group, whereas less than 20% of the JNK inhibitor-negative group achieved normoglycemia. These findings suggest that the JNK pathway is the major mediator of islet deterioration during/immediately after isolation and that JNK inhibition before islet isolation could improve outcomes after pancreatic islet transplantation.