PI3 Kinase and not p42/p44 Appears to be Implicated in the Protection Conferred by Ischemic Preconditioning

M. M. Mocanu, R. M. Bell and D. M. Yellon. PI3 Kinase and not p42/p44 Appears to be Implicated in the Protection Conferred by Ischemic Preconditioning. Journal of Molecular and Cellular Cardiology (2002) 34, 661–668. Ischemic preconditioning results in an immediate phase of protection against lethal ischemia/reperfusion injury that is comprised of both irreversible necrosis and programmed cell death, apoptosis. We hypothesized that preconditioning may activate putative anti-apoptotic pathways, through the induction of either phosphatidyl inositol 3-OH kinase (PI3 kinase) or p42/p44 extracellular receptor kinase, attenuating total cell death. Isolated perfused rat hearts were preconditioned with two cycles of 5 min ischemia and 10 min reperfusion. Then they were frozen for Western blot analysis or subjected to 35 min regional ischemia and 120 min reperfusion prior to infarct size assessment. Selective PI3 kinase inhibitors, wortmannin (W, 100 n M) and LY294002 (LY, 15 μM) and the p42/p44 inhibitor, PD 98059 (PD, 10 and 50 μM), were individually infused during the preconditioning protocol. One further group of hearts received both inhibitors (W and PD). The results were expressed as percentage of infarction within the risk zone. Inhibition of PI3 kinase by either W or LYpartially abrogated the infarct sparing effect of ischemic preconditioning (I/R%: 44.6±2.7 in C, 17.6±2.0 in IP, vs 32.2±4.2 in W, and 30.9±2.6 in LY, P<0.05). Inhibition of ERK phosphorylation however, had no significant effect upon infarct size reduction (17.6±2.0 in ischemic preconditioning vs 21.4±3.0 in IP+10 μM PD and 15.2±1.4 in IP+50 μM PD, P>0.05). Western blot analysis confirmed that PD abrogated the phosphorylation of p42/p44 and LY the phosphorylation of AKT. Combined inhibition with PD+W failed to further attenuate protection (27.6±1.3%,P >0.1). These data appear to demonstrate that the PI3 kinase, but not the p42/p44 cascade, is implicated in early ischemic preconditioning. [Copyright &y& Elsevier]