Вид документа : Статья из журнала Шифр издания : Автор(ы) : Danylovych H. V. Заглавие : Evaluation of functioning of mitochondrial electron transport chain with NADH and FAD autofluorescence Параллельн. заглавия :Оцінка функціонування електронтранспортувального ланцюга мітохондрій міометрія за автофлуоресценцією NADH та FAD Место публикации : Український біохімічний журнал. - 2016. - Т. 88, № 1. - С. 31-43 (Шифр УУ1/2016/88/1) Примечания : Бібліогр.: в кінці ст. MeSH-главная: МЕМБРАННОГО ТРАНСПОРТА БЕЛКИ МИТОХОНДРИАЛЬНЫЕ -- MITOCHONDRIAL MEMBRANE TRANSPORT PROTEINS МОДЕЛИ НА ЖИВОТНЫХ -- MODELS, ANIMAL МИОМЕТРИЙ -- MYOMETRIUM НАД -- NAD ФЛАВИНАДЕНИНДИНУКЛЕОТИД -- FLAVIN-ADENINE DINUCLEOTIDE Аннотация: We prove the feasibility of evaluation of mitochondrial electron transport chain function in isolated mitochondria of smooth muscle cells of rats from uterus using fluorescence of NADH and FAD coenzymes. We found the inversely directed changes in FAD and NADHfluorescence intensity under normal functioning of mitochondrial electron transport chain. The targeted effect of inhibitors of complex I, III and IV changed fluorescence of adenine nucleotides. Rotenone (5 μΜ) induced rapid increase in NADH fluorescence due to inhibition of complex I, without changing in dynamics of FAD fluorescence increase. Antimycin A, a complex III inhibitor, in concentration of 1 μg/ml caused sharp increase in NADH fluorescence and moderate increase in FAD fluorescence in comparison to control. NaN3 (5 mM), a complex IV inhibitor, and CCCP (10 μΜ), a protonophore, caused decrease in NADH and FAD fluorescence. Moreover, all the inhibitors caused mitochondria swelling. NO donors, e.g. 0.1 mM sodium nitroprusside and sodium nitrite similarly to the effects of sodium azide. Energy-dependent Ca2+ accumulation in mitochondrial matrix (in presence of oxidation substrates and Mg-ATP2- complex) is associated with pronounced drop in NADH and FAD fluorescence followed by increasedfluorescence of adenine nucleotides, which may be primarily due to Ca2+- dependent activation of dehydrogenases of citric acid cycle. Therefore, the fluorescent signal of FAD and NADH indicates changes in oxidation state of these nucleotides in isolated mitochondria, which may be used to assay the potential of effectors of electron transport chain