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Modified Glucose as a Sensor to Track the Metabolism of Individual Living Endothelial Cells - Observation of the 'Raman Spectroscopic Signature of Life'

2022; RELX Group (Netherlands); Linguagem: Inglês

10.2139/ssrn.4303613

1556-5068

Anna Pieczara, Aleksandra Borek-Dorosz, Szymon Buda, William J. Tipping, Duncan Graham, Robert Pawłowski, Jacek Młynarski, Małgorzata Barańska,

Spectroscopy Techniques in Biomedical and Chemical Research

Endothelial cells (ECs) play a significant role in a healthy and dysfunctional state, the latter is correlated with a range of lifestyle diseases, such as cardiovascular disorders or some cancers. Therefore, knowledge of the dysfunctional state, cellular metabolism, and especially glucose metabolism, is important from the point of view of studying disease mechanisms. The metabolism and glucose uptake may reflect the physiopathological conditions and cell activity correlated with energy metabolism. Our method of choice for studying changes at the subcellular level is Raman microscopy to investigate the metabolism and tracking of glucose uptake. The study was focused on a modified glucose analogue, 3-O-propargyl-d-glucose (3-OPG), which gives a characteristic and intense Raman band at 2124 cm-1 in both normal and inflamed ECs. So, the alkyne-labeled glucose, 3-OPG, was used as a sensor to track its accumulation and then metabolism in live and fixed ECs by employing two spectroscopic techniques, i.e. spontaneous Raman and stimulated Raman scattering microscopies. The results indicate that 3-OPG is suitable as a sensor for fixed and live cells in normal as well as in inflammatory conditions to follow the glucose metabolism, by analyzing the Raman band of 1602 cm-1 attributed to the glucose metabolites, and the 2124 cm-1 Raman band of glucose accumulation. Additionally, we have shown that glucose metabolism and its uptake are slowed down in inflammation. Gaining further knowledge about metabolic patterns in the endothelium, and especially in the various pathologies affecting ECs, should open up valuable opportunities for entirely new medical treatments.