No confirmation for a causal role of volume-regulated chloride channels in ischemic preconditioning in rabbits.
Volume-regulated chloride channels have recently been proposed to be end-effectors in ischemic preconditioning. The present study attempted to confirm this hypothesis by looking both at cardioprotection and channel activity. In isolated rabbit cardiomyocytes, hypo-osmotic stress (167 mosm/l) induced a current with a magnitude of 2-5 pA/pF at 60 mV. That current could be blocked by the selective chloride channel blockers 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) or indanyloxyacetic acid 94 (IAA-94), but only at 100 microM and 1 m M respectively. Lower concentrations were not effective. Because the channel-blocking concentrations were toxic in isolated perfused rabbit hearts, as evidenced by cessation of cardiac contraction and massive infarction, neither agent could be tested against preconditioning's anti-infarct effect. NPPB and IAA-94 at 1 microM and 10 microM, respectively (the doses used in a previous report), did not affect coronary flow, heart rate and developed pressure, and also did not prevent the infarct size reduction of ischemic preconditioning with 5 min global ischemia/10 min reperfusion preceding 30 min of regional ischemia and 120 min of reperfusion [11. 4(+/-3.6) and (11.1(+/-3.7)% infarction of risk area, respectively]. The volume-regulated chloride and organic osmolyte channel blocker 4, 4;-diisothiocyanostilbene-2,2;-disulfonic acid (DIDS) at 100 microM blocked the hypo-osmotically induced current in myocytes, but again could not be used, since it induced total cessation of cardiac contraction and reduced infarct size in non-preconditioned hearts. Our data do not confirm a prior study on a causal role for volume-regulated chloride channels in the protection of ischemic preconditioning. This hypothesis remains to be adequately tested.
Heusch
G
G
Department of Physiology, University of South Alabama College of Medicine, Mobile, AL 36688, USA.
Liu
G S
GS
Rose
J
J
Cohen
M V
MV
Downey
J M
JM
D000818:Animals; D002478:Cells, Cultured; D018118:Chloride Channels / Q000502:physiology*; D004594:Electrophysiology; D006352:Heart Ventricles / Q000378:metabolism; D006439:Hemodynamics; D019194:Ischemic Preconditioning; D009203:Myocardial Infarction / Q000378:metabolism; D009206:Myocardium / Q000166:cytology; D011817:Rabbits
D018118:Chloride Channels