Effect of lemakalim on action potentials, intracellular calcium, and contraction in guinea pig and human cardiac myocytes.
OBJECTIVE
The aim was to investigate the effects of lemakalim on action potential duration, intracellular free calcium ([free Ca2+]i), and cell contraction in human and guinea pig cardiac myocytes. In addition, the possible modulation by pH of lemakalim induced activation of ATP sensitive potassium (KATP) channels was assessed.


METHODS
Single ventricular myocytes were enzymatically dissociated from adult male guinea pigs (300-600 g). Single myocytes were isolated from human ventricular tissues. Cells were loaded with the acetoxymethyl ester form of fura-2 to monitor changes in [free Ca2+]i and subjected to conventional electrophysiological techniques.


RESULTS
In guinea pig cells, lemakalim (3, 10, 30 microM) reduced action potential duration in a concentration dependent manner. This decrease was accompanied by hyperpolarisation of the resting membrane potential. Lemakalim (3, 10, 30 microM) reduced the systolic fura-2 fluorescence ratio without having a significant effect on diastolic fluorescence and also reduced the cell contraction in concentration dependent manner. Glibenclamide (1 microM), a specific inhibitor of KATP channels, did not affect action potential duration, fura-2 fluorescence ratio, or cell contraction in the absence of lemakalim. However, the same dose of glibenclamide markedly inhibited the lemakalim induced decrease in action potential duration, fura-2 fluorescence ratio, and cell contraction. Reducing extracellular pH enhanced the decrease in action potential duration induced by lemakalim. In human ventricular myocytes, lemakalim (3, 10 and 30 microM) caused a decrease in action potential duration and systolic fura-2 fluorescence ratio. The reduction in action potential duration and fura-2 fluorescence ratio was also reversed by glibenclamide (1 microM).


CONCLUSIONS
These results suggest that lemakalim reduces systolic [free Ca2+]i by activating ATP sensitive potassium channels which results in a decrease of action potential duration in guinea pig and human ventricular myocytes. The reduction in [free Ca2+]i mediates the negative inotropic effect induced by lemakalim. In addition, pH may modulate the KATP channel activation by the channel opener.
Jiang
C
C
Department of Cardiac Medicine, National Heart and Lung Institute, London, United Kingdom.
Mochizuki
S
S
Poole-Wilson
P A
PA
Harding
S E
SE
MacLeod
K T
KT
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D001578:Benzopyrans / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D019806:Cromakalim; D004305:Dose-Response Relationship, Drug; D016257:Fura-2; D005905:Glyburide / Q000494:pharmacology; D006168:Guinea Pigs; D006321:Heart / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D011758:Pyrroles / Q000494:pharmacology*; D014665:Vasodilator Agents / Q000494:pharmacology*
D001578:Benzopyrans; D011758:Pyrroles; D014665:Vasodilator Agents; D019806:Cromakalim; D005905:Glyburide; D002118:Calcium; D016257:Fura-2