ИСТИНА

The ATP amount and rate of ATP production by human mitochondria are decreased in presence of Ab(1–42) peptide, which is known that involved in pathogenesis of Alzheimer’s disease. The1st experimental group of SH-SY5Y human neuroblastoma cell line were cultivated 24h (~90% confluence) with 200 nM Ab monomerased by standard procedure (Jao 1997, Dzinic 2018),2nd group (control) of cells had no additives, and the 3d group contained 0.1% DMSO (this amount of DMSO were added to the 1st group with Ab). Mitochondria were isolated by standard procedure (Martin 1998, Daum 1982). To isolated mitochondria were added substrates of Complex I, II and IV and corresponded inhibitors of OxPhos Complexes – in separate experiments to highlight the changes of activity of each Complex induced by Ab. Immediately before the measurements of luminescence to mitochondria were added ADP and luciferase-luciferin mixture (Ugarova et al. Patent RU 2420594, 2011). The luminescence (RLU) depending on time was plotted for each experimental group. The maximum values of the first derivatives of the left sides of the obtained bell shaped curves (the rise of the ATP production) were compared (the rate of ATP production). Also were compared the the peak heights (corresponded to the amount of the produced ATP). The presence of DMSO led to the slight lowering of ATP production by human mitochondria – amount on 9.3% and rate on 6.6% if compare with the control group(mean value for the Complexes I, II, IV). Cultivation of cells with Ab makes mitochondria to produce 26.3% lower ATP (if compare with the group treated with DMSO) with 48.3% lower rate. Widely used the methods which include the disruption of cells or mitochondria to measure the total amount of ATP molecules in the lysates (Drew 2003, Gao 2009, Lim 2011). But we suppose that the plotting the level of luminescence (ATPconcentration) in time can give more information.