Вплив флокаліну на розвиток апоптозу та некрозу при аноксії–реоксигенації ізольованих неонатальних кардіоміоцитів

Abstract

The influence of the new cardioprotector flocalin was investigated on the culture of rat's neonatal cardiomyocytes during anoxia-reoxygenation modelling. The mechanisms of apoptosis and necrosis were investigated under influence of ATP-sensitive potassium (K(ATP)) channels activation and in conditions of blocking of the L-type calcium (VGCCs) channels. Flocalin was added in the culture medium in the dose 5 and 20 microM at 2 minutes before anoxia (30 minutes) and following reoxygenation (60 minutes). These doses are near to: the first dose means the opening of K(ATP) channels and the second one means the IC50 block of VGCCs. It is discovered that in dose 5 microM of flocalin drew the change of correlation of living, necrotizing and apoptizing cells drew side-shifting living. The number of live cells was almost the same like in control (experiments without anoxia-reoxygenation modelling). The opening of K(ATP) channels decreases necrosis in two times and fully prevented development of apoptosis which was induced anoxia-reoxygenation modelling. Flocalin depressed the apoptosis of neonatal cardiomyocytes so that he was on to 36% less than in control group (without anoxia-reoxygenation). But in the high dose (20 microM) that provokes not only K(ATP) channels opening but also IC50 block of VGCCs cardioprotection was not detected after modelling of anoxia-reoxygenation. The last can be investigation both enough strong activating of the potassium channels and by investigation of both factors are opening of potassium and inhibition of VGCCs channels and, accordingly, substantial diminishing of level of introcellular Ca2+ and violation of metabolic processes yet to anoxia-reoxygenation. Thus, small doses of flocalin, that induce moderate opening of K(ATP) channels significantly decrease the number of necrotic and apoptotic cells in culture of rat neonatal cardiomyocytes induced by anoxia-reoxygenation.