Abstract 2321: Reactive Oxygen Species-Induced Activation of 5′AMP-Activated Protein Kinase Mediates Cardioprotection Against Ischemia and Reperfusion Injury
The myocardium possesses intrinsic protective mechanisms against ischemia and reperfusion injury (I/R). 5′AMP-activated protein kinase (AMPK) is known as regulator of cellular energy status and is reduced during diabetes mellitus. Recently, AMPK is also implicated in ischemic preconditioning leading to cardioprotection against I/R. We hypothesize that AMPK is involved in anesthetic-induced cardioprotection and that this AMPK activation is evoked by production of reactive oxygen species (ROS). Isolated Langendorff-perfused rat hearts were subjected to 35 minutes of global ischemia followed by 120 minutes of reperfusion. Hearts were divided into 2 groups: a Control group and a Sevo group receiving three times 5-minute episodes of 2.5 vol% sevoflurane before I/R. AMPK phosphorylation was determined with Western Blot analysis. Cardioprotection was assessed as recovery of left ventricular pressure after I/R and as infarct size using triphenyltetrazolium chloride staining. In the Control group, I/R resulted in a twofold increase in phosphorylation levels of AMPK (Control 1.0 ± 0.1 vs. Control-I/R: 2.3 ± 0.1 a.u., n = 4, p < 0.05). Sevoflurane preconditioning did not immediately, prior to ischemia, affect the AMPK phosphorylation (Sevo 0.9 ± 0.2 vs. Control 1.0 ± 0.2, n = 6), but doubled the increase in AMPK phosphorylation to control after ischemia (Sevo-I: 2.0 ± 0.5 (vs. Control-I), n = 6, p < 0.05), as well as after I/R (Sevo-I/R: 2.1 ± 0.3 vs. (Control-I/R), n = 6, p < 0.05). The AMPK-inhibitor compound C (1 and 10 μM) reduced the increase in AMPK phosphorylation and abolished the cardioprotection derived from functional recovery and infarct size. In addition, the ROS-scavenger n-(2-mercaptopropionyl)-glycine (MPG, 1mM) also reduced the sevoflurane-mediated increase in AMPK phosphorylation and completely prevented cardioprotection. These results demonstrate for the first time a direct link between AMPK activation and the production of ROS in cardioprotection. We conclude that anesthetic-induced AMPK activation protects the heart against I/R and relies on production of ROS, which might be especially important in the context of impaired cardioprotection in the diabetic myocardium.