Direct effects of K(ATP) channel openers pinacidil and diazoxide on oxidative phosphorylation of mitochondria in situ.
K(ATP) channel openers protect ischemic-reperfused myocardium by mimicking ischemic preconditioning, however, the protection mechanisms have not been fully clarified yet. Since the skinned fibers technique gives an opportunity to investigate an entire population of mitochondria in their native milieu, in this study we have investigated the effects of K(ATP) channel openers pinacidil and diazoxide on the respiration rate of rat heart mitochondria in situ, oxidizing physiological substrates pyruvate and malate (6+6 mM). Respiration rates were recorded by the means of Clark-type oxygen electrode in the physiological salt solution (37 degrees C). Our results showed that both pinacidil and diazoxide (60-1250 muM) in a concentration-dependent manner increased pyruvate-malate supported State 2 respiration rate of skinned cardiac fibers (59.1 +/- 5.1 nmol O/min/mg fiber dry weight, RCI 2.6 +/- 0.2, n=4) by 15-120%. Moreover, diazoxide did not affect, whereas pinacidil (60-1250 muM) decreased the State 3 respiration rate of skinned cardiac fibers (116.6 +/- 13.6 nmol O/min/mg fiber dry weight, RCI 2.3 +/- 0.2, n=4) by 4-27%. Thus, common effect for both K(ATP) channel openers is uncoupling of pyruvate and malate oxidizing mitochondria in skinned cardiac fibers, whereas pinacidil under same conditions also inhibits mitochondrial respiratory chain. Since mitochondria in situ resemble to the great extent mitochondria in vivo, our results suggest that uncoupling and/or respiratory chain inhibition could play a role in the cardioprotection by K(ATP) channel openers.
Kopustinskiene
Dalia M
DM
Laboratory of Biochemistry, Institute for Biomedical Research of Kaunas University of Medicine, 50009, Kaunas, Lithuania. daliammk@mail.lt
Liobikas
Julius
J
Skemiene
Kristina
K
Malinauskas
Faustas
F
Toleikis
Adolfas
A
D000818:Animals; D003981:Diazoxide / Q000494:pharmacology*; D054086:KATP Channels / Q000378:metabolism*; D008293:Malates / Q000378:metabolism; D008297:Male; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism; D010085:Oxidative Phosphorylation; D020110:Pinacidil / Q000494:pharmacology*; D019289:Pyruvic Acid / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D056152:Respiratory Rate; D014665:Vasodilator Agents / Q000494:pharmacology*
D054086:KATP Channels; D008293:Malates; D014665:Vasodilator Agents; D020110:Pinacidil; C030298:malic acid; D019289:Pyruvic Acid; D003981:Diazoxide