Strutyns'kyĭ R B, Nagibin V S, Strutyns'ka N A, Ianchiĭ O R, Moĭbenko O O
Fiziol Zh (1994). 2013;59(3):3-9.
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.
在缺氧-复氧模型中，研究了新型心脏保护剂氟卡林对大鼠新生心肌细胞培养的影响。在ATP敏感性钾（K(ATP)）通道激活以及L型钙（VGCCs）通道阻断的情况下，研究了细胞凋亡和坏死的机制。在缺氧（30分钟）前2分钟以及复氧（60分钟）后，将氟卡林以5和20微摩尔的剂量添加到培养基中。这些剂量接近：第一个剂量意味着K(ATP)通道开放，第二个剂量意味着VGCCs的IC50阻断。发现5微摩尔剂量的氟卡林使存活、坏死和凋亡细胞的相关性发生变化，导致存活细胞出现侧移。存活细胞的数量与对照组（无缺氧-复氧模型的实验）几乎相同。K(ATP)通道的开放使坏死减少了两倍，并完全阻止了由缺氧-复氧模型诱导的细胞凋亡的发生。氟卡林抑制了新生心肌细胞的凋亡，使其比对照组（无缺氧-复氧）减少了36%。但在高剂量（20微摩尔）下，不仅会引发K(ATP)通道开放，还会导致VGCCs的IC50阻断，在缺氧-复氧建模后未检测到心脏保护作用。最后这一点可以通过对钾通道的足够强激活以及对钾通道开放和VGCCs通道抑制这两个因素的研究来进行探究，相应地，细胞内Ca2+水平会大幅降低，并且在缺氧-复氧之前代谢过程就会受到破坏。因此，小剂量的氟卡林，即诱导K(ATP)通道适度开放，可显著减少缺氧-复氧诱导的大鼠新生心肌细胞培养物中坏死和凋亡细胞的数量。
