Glycolysis process activation in preserved red blood cells by nanotechnological treatment of resuspending solutions

  • A. N. Belousov Laboratory of Applied Nanotechnology of Belousov
  • E. I. Malygon Kharkiv National Medical University
  • T. O. Kalynychenko Institute of Hematology and Transfusiology of the National Academy of Medical Sciences of Ukraine
  • E. Y. Belousova Kharkiv National Medical University
  • V. V. Yavorskiy Kharkiv National Medical University
  • M. Y. Anoshyna Institute of Hematology and Transfusiology of the National Academy of Medical Sciences of Ukraine
Keywords: magnetite nanoparticles; resuspending solution; preserved erythrocytes; membrane-protective effect; anaerobic glycolysis; pentose cycle.


Currently, the use of nanotechnology opens up new opportunities to influence the processes of anaerobic glycolysis and the activity of hexose monophosphate reactions in preserved erythrocytes. Components containing donor red blood cells on CPDA-1 preservative were examined. Modified solutions of 0.9% NaCl and with 5% glucose were used as resuspending solutions. The solutions were treated with magnetite nanoparticles (ICNB brand) by the Belousov method. The amounts of 2,3-DPG, ATP, reduced glutathione, and glutathione peroxidase were determined by spectrophotometry. This study opens up new possibilities for increasing the shelf life and functional activity of preserved erythrocytes. The study showed a reliable increase in ATP and reduced glutathione, a decrease in 2,3-DPG and glutathione peroxidase. It was found that the activation of anaerobic glycolysis was less pronounced in tests with modified physiological saline than in tests with glucose solution. On the contrary, the pentose glucose oxidation cycle prevailed. A comprehensive analysis of the data obtained indicates the membrane-protective effect of the modified resuspending solutions. The membrane-protective effect is due to an increase in ATP and reduced glutathione, which ensures the redox potential of the cell in an equilibrium state. Magnetite nanoparticles (ICNB) change the mobility and orientation of hydrogen protons in resuspending solutions. This polarizes the aqueous sector of the erythrocyte microenvironment due to van der Waals forces, which is the main reason for activation of ATP phosphate residue hydrolysis and switching of intracellular enzymes regulating anaerobic glycolysis and pentose phosphate cycle into the active state. As a result, transmembrane metabolism and metabolism change, the energy state of erythrocytes changes, and enzymes are activated. All this has a significant impact on the energy supply of preserved red blood cells and preservation of their functional activity under storage conditions at 2 to 6 ºС.


Agarwal, V., Gupta, V., Bhardwaj, V. K., Singh, K., Khullar, P., & Bakshi, M. S. (2022). Hemolytic response of iron oxide magnetic nanoparticles at the interface and in bulk: Extraction of blood cells by magnetic nanoparticles. ACS Applied Materials and Interfaces, 14(5), 6428–6441.

Belousov, A. N. (2011). The use of magnetite nanoparticles in applied medicine. Materials Science Forum, 694, 205–208.

Belousov, A. N. (2013). Myth and reality application of magnetite nanoparticles as selective contrasting means of the malignant tumors in MRI investigation. Biomedical Engineering Research, 2(3), 147–152.

Belousov, A. N. (2014). The role of magnetite nanoparticles (ICNB) in discovery new factor which influence on permeability of erythrocytes and eryptosis. Open Access Library Journal, 1, e1055.

Belousov, A. N., Malygon, E., Yavorskiy, V., & Belousova, E. (2018). Application of the standardized form magnetite nanoparticles (ICNB) in creature simple and practical method of additive modernization of preservation solutions for red blood cells. Global Journal of Anesthesia and Pain Medicine, 1(1), 1.

Belousov, A., Kalynychenko, T., Malygon, E., Anoshyna, M., Yagovdik, M., Yavorskiy, V., & Belousova, E. (2021). Study of effects a new resuspending solution which was nanotechnologically upgraded on lipoperoxidation, catalase activity and red blood cell peroxidation resistance in donor blood components. Archives in Biomedical Engineering and Biotechnology, 6(2), 1–6.

Belousov, A., Malygon, E., Yavorskiy, V., & Belousova, E. (2019). Innovative method of nanotechnology to increase the storage time of RBCs due by stabilizing the molecular structure of proteins and lipids of erythrocyte membranes. Biomedical Journal of Scientific and Technical Research, 13(4), 10079–10087.

Benesch, R., & Benesch, R. E. (1967). The effect of organic phosphates from the human erythrocyte on the allosteric properties of hemoglobin. Biochemical and Biophysical Research Communications, 26(2), 162–167.

Brewer, G. J. (1974). 2,3-DPG and erythrocyte oxygen affinity. Annual Review of Medicine, 25, 29–38.

Devasagayam, T. P. A., Tilak, J. C., Boloor, K. K., Sane, K. S., Ghaskadbi, S. S., & Lele, R. D. (2004). Free radicals and antioxidants in human health: current status and future prospects. The Journal of the Association of Physicians of India, 52, 794–804.

Eschenko, N. D., & Volski, G. G. (Eds.) (1982). Metody biokhimicheskikh issledovanij [Methods of biochemical studies]. Leningradskij Gosudarstvennyj Universitet, Leningrad (in Russian).

Francis, R. O., D’Alessandro, A., Eisenberger, A., Soffing, M., Yeh, R., Coronel, E., Sheikh, A., Rapido, F., Carpia, F. L., Reisz, J. A., Gehrke, S., Nemkov, T., Thomas, T., Schwartz, J., Divgi, C., Kessler, D., Shaz, B. H., Ginzburg, Y., Zimring, J. C., Spitalnik, S. L., & Hod, E. A. (2020). Donor glucose-6-phosphate dehydrogenase deficiency decreases blood quality for transfusion. Journal Clinical Investigation, 130(5), 2270–2285.

Guitton, J., Servanin, S., & Francina, A. (2003). Hexose monophosphate shunt activities in human erythrocytes during oxidative damage induced by hydrogen peroxide. Archive Toxicology, 77(7), 410–417.

Halprin, K. M., & Ohkawara, A. (1967). The measurement of glutathione in human epidermis using glutathione reductase. The Journal of Investigative Dermatology, 48(2), 149–152.

Hayyan, M., Hashim, M. A., & Al Nashef, I. M. (2016). Superoxide ion: Generation and chemical implications. Chemical Reviews, 116(5), 3029–3085.

Jacobasch, G., Bleiber, R., & Schönian, G. (1982). Metabolism of the hexose monophosphate shunt in glucose-6-phosphate dehydrogenase deficiency and closely interrelated reactions. Haematologia, 15(4), 401–407.

Lu Shelly, C. (2013). Glutathione synthesis. Biochimica et Biophysica Acta – General Subjects, 1830(5), 3143–3153.

McMahon, T. J., Darrow, C. C., Brooke, A. H., & Zhu, H. (2021). Generation and export of red blood cell ATP in health and disease. Frontiers in Physiology, Red Blood Cell Physiology, 5(12), 754638.

Melkonian, E. A., & Schury, M. P. (2022). Biochemistry, anaerobic glycolysis. StatPearls Publishing, Treasure Island.

Mills, B. J., Richie, J. P., & Lang, C. A. (1994). Glutathione disulfide variability in normal human blood. Analytical Biochemistry, 222(1), 95–101.

Moin, V. M. (1986). A simple and specific method for determining glutathione peroxidase activity in erythrocytes. Laboratornoe Delo, 12, 724–727 (in Russian).

Mranova, I. S. (1975). Determination of 2,3DPG and ATP in red blood cells. Laboratory Business, 7, 652–654.

Murakami, K., Kondo, T., Ohtsuka, Y., Fujiwara, Y., Shimada, M., & Kawakami, Y. (1989). Impairment of glutathione metabolism in erythrocytes from patients with diabetes mellitus. Metabolism Clinical and Experimental, 38(8), 753–758.

Novello, F., & McLean, P. (1968). The pentose phosphate pathway of glucose metabolism. Measurement of the non-oxidative reactions of the cycle. The Biochemical Journal, 107(6), 775–791.

Pastore, A., Piemonte, F., Locatelli, M., Lo Russo, A., Gaeta, L. M., Tozzi, G., & Federici, G. (2001). Determination of blood total, reduced, and oxidized glutathione in pediatric subjects. Clinical Chemistry, 47(8), 1467–1469.

Pietkiewicz, J., Danielewicz, R., Wandzel, C., Beznosiuk, J., Szuba, A., Samsel-Czekała, M., & Gamian, A. (2021). Influence of water polarization caused by phonon resonance on catalytic activity of enolase. Journal of the American Chemical Society, 6, 4255–4261.

Pompella, A., Visvikis, A., Paolicchi, A., De Tata, V., & Casini, A. F. (2003). The changing faces of glutathione, a cellular protagonist. Biochemical Pharmacology, 66(8), 1499–1503.

Richardson, S. R., & O’Malley, G. F. (2022). Glucose 6 phosphate dehydrogenase deficiency. StatPearls Publishing, Treasure Island.

Richie Jr., J. P., Abraham, P., & Leutzinger, Y. (1996). Long-term stability of blood glutathione and cysteine in humans. Clinical Chemistry, 42(7), 1100–1105.

Schafer, F. Q., & Buettner, G. R. (2001). Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radical Biology and Medicine, 30(11), 1191–1212.

Scholz, R. W., Graham, K. S., Gumpricht, E., & Reddy, C. C. (1989). Mechanism of interaction of vitamin E and glutathione in the protection against membrane lipid peroxidation. Annals of the New York Academy of Sciences, 570(1), 514–517.

Stanton, R. C. (2012). Glucose-6-phosphate dehydrogenase, NADPH, and cell survival. International Union of Biochemistry and Molecular Biology Life, 64(5), 362–369.

Turrens, J. F. (2003). Mitochondrial formation of reactive oxygen species. The Journal of Physiology, 552(2), 335–344.

Van't Erve, T. J., Wagner, B. A., Ryckman, K. K., Raife, T. J., & Buettner, G. R. (2013). The concentration of glutathione in human erythrocytes is a heritable trait. Free Radical Biology and Medicine, 65, 742–749.

Waszczak, C., Carmody, M., & Kangasjärvi, J. (2018). Reactive oxygen species in plant signaling. Annual Review of Plant Biology, 69, 209–236.

Yoshida, T., Prudent, M., & D’Alessandro, A. (2019). Red blood cell storage lesion: Causes and potential clinical consequences. Blood Transfusion, 17(1), 27–52.

How to Cite
Belousov, A. N., Malygon, E. I., Kalynychenko, T. O., Belousova, E. Y., Yavorskiy, V. V., & Anoshyna, M. Y. (2023). Glycolysis process activation in preserved red blood cells by nanotechnological treatment of resuspending solutions . Regulatory Mechanisms in Biosystems, 14(1), 10-15.