Pathological cells determination based on blue autofluorescence
Pathological cells have a higher level of autofluorescence in the blue spectrum than control cells that can be potentially used for pathology identification. In this work, using inhibitory analysis we identify the intracellular sources of autofluorescence in pathological cells.
The cell lines of 20-day human fibroblast cultures of control patients were used. The fluorescence was recorded using a ZEISS LSM 900 confocal microscope with the Airyscan 2 system (Carl Zeiss AG, Germany) with excitation at 455 nm. For assessment of the role of FAD MAO in total autofluorescence, the baseline level of endogenous autofluorescence was initially recorded for 2 minutes, then adrenaline (10 μM) and selegiline (20 μM) were added. MAO was activated for the oxidative deamination of adrenaline, the coenzyme of which is autofluorescent FAD. Thus, the maximum values of autofluorescence were recorded. Selegiline is an irreversible MAO A and B inhibitor led to a minimum level of autofluorescence in this enzyme.
The statistical analysis of the results was performed with the preliminary division of cells into two groups – with high autofluorescence (presumably aging and pathological) and with low autofluorescence. The MAO pool was 10.065% ± 2.10095 for cells with high autofluorescence, which is more than for cells with low fluorescence. This parameter was 6.814% ± 0.93281 for the last ones. The statistically significant difference is p<=0.0002. In pathological cells with high fluorescence, MAO is actively working, which leads to an excessive amount of aldehydes that are formed as a product of the adrenaline deamination. These substances inhibit the work of succinate dehydrogenase that forms a non-fluorescent FAD from fluorescent FADH2. Thus, the fluorescence in pathological cells is higher than in conditionally healthy and young cells due to the failure of the complex II of the electron transport chain.
Thus, it is possible to determine the state of a certain cell by knowing the MAO pool and evaluating the efficiency of the complex II of the electron transport chain. This approach can be used as a timely diagnosis and identification of any pathological processes.
This study was supported by the grant from the Russian Federation Government No. 075-15-2019-1877.
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Orel State University named after I.S. Turgenev
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