title: Role of PI3Kα and sarcolemmal ATP-sensitive potassium channels in epoxyeicosatrienoic acid mediated cardioprotection.
abstract: OBJECTIVE
  Epoxyeicosatrienoic acids (EETs) are cytochrome P450 epoxygenase metabolites of arachidonic acid that have known cardioprotective properties. While the mechanism(s) remains unknown, evidence suggests that phosphoinositide 3-kinase (PI3K) and sarcolemmal ATP-sensitive potassium channels (pmK(ATP)) are important. However the role of specific PI3K isoforms and corresponding intracellular mechanisms remains unknown.
  
  
  RESULTS
  To study this, mice hearts were perfused in Langendorff mode for 40 min of baseline and subjected to 20 or 30 min of global no-flow ischemia followed by 40 min of reperfusion. C57BL6 mice perfused with 11,12-EET (1 μM) had improved postischemic recovery, whereas co-perfusion with PI3Kα inhibitor, PI-103 (0.1 μM), abolished the EET-mediated effect. In contrast, blocking of PI3Kβ or PI3Kγ isoforms failed to inhibit EET-mediated cardioprotection. In addition to the improved post-ischemic recovery, increased levels of p-Akt, decreased calcineurin activity and decreased translocation of proapoptotic protein BAD to mitochondria were noted in EET-treated hearts. Perfusion of 11,12-EET to Kir6.2 deficient mice (pmK(ATP)) failed to improve postischemic recovery, decrease calcineurin activity and translocation of proapoptotic protein BAD, however increased levels of p-Akt were still observed. Patch-clamp experiments demonstrated that 11,12-EET could not activate pmK(ATP) currents in myocytes pre-treated with PI-103. Mechanistic studies in H9c2 cells demonstrate that 11,12-EET limits anoxia-reoxygenation triggered Ca(2+) accumulation and maintains mitochondrial ΔΨm compared to controls. Both PI-103 and glibenclamide (10 μM, pmK(ATP) inhibitor) abolished EET cytoprotection.
  
  
  CONCLUSIONS
  Together our data suggest that EET-mediated cardioprotection involves activation of PI3Kα, upstream of pmK(ATP), which prevents Ca(2+) overload and maintains mitochondrial function.
authors.affiliation: Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, AB, Canada.
authors.forename: Sri N
authors.initials: SN
authors.lastname: Batchu
authors.forename: Ketul R
authors.initials: KR
authors.lastname: Chaudhary
authors.forename: Haitham
authors.initials: H
authors.lastname: El-Sikhry
authors.forename: Wei
authors.initials: W
authors.lastname: Yang
authors.forename: Peter E
authors.initials: PE
authors.lastname: Light
authors.forename: Gavin Y
authors.initials: GY
authors.lastname: Oudit
authors.forename: John M
authors.initials: JM
authors.lastname: Seubert
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002316:Cardiotonic Agents / Q000494:pharmacology*; D002460:Cell Line; D058543:Class Ia Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D006321:Heart / Q000187:drug effects*; D000860:Hypoxia; D007527:Isoenzymes / Q000378:metabolism; D054086:KATP Channels / Q000378:metabolism*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015428:Myocardial Reperfusion Injury / Q000378:metabolism / Q000503:physiopathology / Q000517:prevention & control; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism*; D012508:Sarcolemma / Q000201:enzymology / Q000378:metabolism*
chemical_list: D002316:Cardiotonic Agents; D007527:Isoenzymes; D054086:KATP Channels; C046783:11,12-epoxy-5,8,14-eicosatrienoic acid; D058543:Class Ia Phosphatidylinositol 3-Kinase; D015126:8,11,14-Eicosatrienoic Acid; D002118:Calcium