Ischemic preconditioning activates phosphatidylinositol-3-kinase upstream of protein kinase C

The present study is designed to test whether phosphatidylinositol 3-kinase (PU-kinase) has a role in the signaling pathway in ischemic preconditioning (PC) and whether it is proximal or distal to protein kinase C (PKC). Before 20 minutes of global ischemia, Langendorff-perfuse rat hearts were perfused for 20 minutes (control); preconditioned with 4 cycles of ti-minute ischemia and 5-minute reflow (PC); treated with either wortmannin (WM) or LY 294002, (LY), each of which is a PU-kinase inhibitor, for 5 minutes before and throughout PC; treated with 1,2-dioctanoyl-sn-glycerol (DOG), an activator of PKC for 10 minutes (DOG); treated identically to the DOG group except with WM added 10 minutes before and during perfusion with DOG; or treated with either WM or LY for 25 minutes. Recovery of left ventricular developed pressure (LVDP; percentage of initial preischemic LVDP), measured after 30 minutes of reflow, was improved by PC (72+/-2% versus 36+/-4% in control; P<0.001), and this was blocked by WM and LY (41+/-4% and 33+/-5%, respectively; P<0.05 compared with PC). DOG addition improved postischemic LVDP (67+/-6%; P<0.001 compared with control), but in contrast to its effect on PC, WM did not completely eliminate the protective effect of DOG (52+/-4%; P>0.05 compared with DOG; P<0.05 compared with control). PC induced phosphorylation of ptotein kinase B and translocation of PKC epsilon, and it increased NO production, and these effects were blocked by WM, which suggests a role for P13-kinase in PC upstream of PKC and NO.