title: Signal transduction of flumazenil-induced preconditioning in myocytes.
abstract: The objective of this study was to examine the role of oxygen radicals, protein kinase C (PKC), and ATP-sensitive K(+) (K(ATP)) channels in mediating flumazenil-produced preconditioning. Chick cardiomyocyte death was quantified using propidium iodide, and oxygen radical generation was assessed using 2',7'-dichlorofluorescin oxidation. Preconditioning was initiated with 10 min of ischemia followed by 10 min of reoxygenation. Alternatively, flumazenil was infused for 10 min and removed 10 min before ischemia. Flumazenil (10 microM) and preconditioning increased oxygen radicals [1,693 +/- 101 (n = 3) and 1,567 +/- 98 (n = 3), respectively, vs. 345 +/- 53 (n = 3) in control] and reduced cell death similarly [22 +/- 3% (n = 5) and 18 +/- 2% (n = 6), respectively, vs. controls 49 +/- 5% (n = 8)]. Protection and increased oxygen radicals by flumazenil were abolished by pretreatment with the antioxidant thiol reductant 2-mercaptopropionyl glycine (800 microM; 52 +/- 10%, n = 6). Specific PKC inhibitors Go-6976 (0.1 microM) and chelerythrine (2 microM), given during ischemia and reoxygenation, blocked flumazenil-produced protection (47 +/- 5%, n = 6). The PKC activator phorbol 12-myristate 13-acetate (0.2 microM), given during ischemia and reoxygenation, reduced cell death similarly to that with flumazenil [17 +/- 4% (n = 6) and 22 +/- 3% (n = 5)]. Finally, 5-hydroxydecanoate (1 mM), a selective mitochondrial K(ATP) channel antagonist given during ischemia and reoxygenation, abolished the protection of flumazenil and phorbol 12-myristate 13-acetate. Thus flumazenil mimics preconditioning to reduce cell death in cardiomyocytes. Oxygen radicals activate mitochondrial K(ATP) channels via PKC during the process.
authors.affiliation: Department of Anesthesia and Critical Care, The University of Chicago, Chicago, Illinois 60637, USA. zyao@airway2.uchicago.edu
authors.forename: Z
authors.initials: Z
authors.lastname: Yao
authors.forename: B C
authors.initials: BC
authors.lastname: McPherson
authors.forename: H
authors.initials: H
authors.lastname: Liu
authors.forename: Z
authors.initials: Z
authors.lastname: Shao
authors.forename: C
authors.initials: C
authors.lastname: Li
authors.forename: Y
authors.initials: Y
authors.lastname: Qin
authors.forename: T L
authors.initials: TL
authors.lastname: Vanden Hoek
authors.forename: L B
authors.initials: LB
authors.lastname: Becker
authors.forename: P T
authors.initials: PT
authors.lastname: Schumacker
mesh_terms: D000818:Animals; D002227:Carbazoles / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002642:Chick Embryo; D003600:Cytosol / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005442:Flumazenil / Q000494:pharmacology*; D018757:GABA Modulators / Q000494:pharmacology*; D006861:Hydrogen Peroxide / Q000378:metabolism; D007211:Indoles / Q000494:pharmacology; D019157:Ischemic Preconditioning, Myocardial; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D017202:Myocardial Ischemia / Q000188:drug therapy / Q000503:physiopathology; D009206:Myocardium / Q000166:cytology*; D015221:Potassium Channels / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D018079:Receptors, GABA / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects*; D008625:Tiopronin / Q000494:pharmacology
chemical_list: D002227:Carbazoles; D004791:Enzyme Inhibitors; D018757:GABA Modulators; D007211:Indoles; D015221:Potassium Channels; D017382:Reactive Oxygen Species; D018079:Receptors, GABA; C081021:Go 6976; D005442:Flumazenil; D006861:Hydrogen Peroxide; D008625:Tiopronin; D011493:Protein Kinase C