title: Chloride channel inhibition blocks the protection of ischemic preconditioning and hypo-osmotic stress in rabbit ventricular myocardium.
abstract: The objective of this study was to examine the role of chloride (Cl-) channels in the myocardial protection of ischemic preconditioning (IP). Isolated rabbit ventricular myocytes were preconditioned with 10-minute simulated ischemia (SI) and 20-minute simulated reperfusion (SR) or not preconditioned (control). The myocytes then received 180-minute SI or 45-minute SI/120-minute SR. Indanyloxyacetic acid 94 (IAA-94, 10 micromol/L) or 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB, 1 micromol/L) was administered before IP or before SI or SI/SR to inhibit Cl- channels. Electrophysiological studies indicate that these drugs, at the concentrations used, selectively abolished Cl- currents activated under hypo-osmotic conditions (215 versus 290 mOsm). IP significantly (P<0.001) reduced the percentage of dead myocytes after 60-minute (30.8+/-1.3%, mean+/-SEM), 90-minute (35.3+/-1.3%), and 120-minute (39.2+/-1.7%) SI compared with controls (44.7+/-1.6%, 54.5+/-1.3%, and 58.9+/-1.8%, respectively) and after 45-minute SI/120-minute SR (36.3+/-0.6%) compared with control (56.6+/-2.2%). Hypo-osmotic stress also produced protection similar to IP. IAA-94 or NPPB abolished the protection of both IP and hypo-osmotic stress. In buffer-perfused rabbit hearts preconditioned with three 5-minute ischemia/10-minute reperfusion cycles given before the 40-minute long ischemia and 60-minute reperfusion, IP significantly (P<0.0001) reduced infarct size (IP+vehicle, 4.7+/-0.9%, versus control+vehicle, 26.6+/-3.3%; mean+/-SEM). Again, IAA-94 or NPPB abolished the protection of IP. Our results implicate Cl- channels in the IP protection of the myocardium against ischemic/reperfusion injury and demonstrate that hypo-osmotic stress is capable of preconditioning cardiomyocytes.
authors: Divisions of Cardiovascular Research and Pathology, Research Institute, The Hospital for Sick Children, Toronto, Ontario, Canada. diazport@sickkids.on.ca
authors: R J
authors: RJ
authors: Diaz
authors: V A
authors: VA
authors: Losito
authors: G D
authors: GD
authors: Mao
authors: M K
authors: MK
authors: Ford
authors: P H
authors: PH
authors: Backx
authors: G J
authors: GJ
authors: Wilson
mesh_terms: D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D002478:Cells, Cultured; D018118:Chloride Channels / Q000037:antagonists & inhibitors* / Q000502:physiology; D004594:Electrophysiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006352:Heart Ventricles / Q000737:chemistry / Q000166:cytology / Q000473:pathology; D007038:Hypotonic Solutions; D019157:Ischemic Preconditioning, Myocardial; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D009203:Myocardial Infarction / Q000378:metabolism / Q000473:pathology; D017202:Myocardial Ischemia / Q000378:metabolism*; D009206:Myocardium / Q000737:chemistry* / Q000166:cytology / Q000473:pathology; D009336:Necrosis; D009997:Osmotic Pressure; D010477:Perfusion; D010617:Phenanthridines / Q000494:pharmacology; D011817:Rabbits
chemical_list: D000470:Alkaloids; D053119:Benzophenanthridines; D018118:Chloride Channels; D004791:Enzyme Inhibitors; D007038:Hypotonic Solutions; D010617:Phenanthridines; C016299:chelerythrine