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Bìol. Tvarin. 2022; 24 (4): 8–11.
https://doi.org/10.15407/animbiol24.04.008
Received 02.05.2022 ▪ Revision 14.09.2022 ▪ Accepted 14.12.2022 ▪ Published online 30.12.2022

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Efficiency of amphiphilic compounds in rabbit erythrocytes posthypertonic shock depending on temperature conditions

O. E. Nipot, N. A. Ershova, N. M. Shpakova, S. S. Ershov, O. O. Shapkina

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Institute for Problems of Cryobiology and Cryomedicine NAS of Ukraine,
23 Pereyaslavska str., Kharkiv, 61016, Ukraine

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The influence of temperature conditions on the level of damage of rabbit erythrocytes under posthypertonic shock and the level of their protection by amphiphilic compounds was investigated. We observed the maximum cell damage at 0°C. When the temperature increased to 20°C, the level of hemolysis decreased by 1.8 times. Further increase in temperature up to 37°C did not lead to a decrease in damage. The investigated amphiphilic compounds at 0°C and 10°C effectively protected rabbit erythrocytes from posthypertonic shock. Reduction of hemolytic damage was 2–3 times. At 20°C amphiphilic compounds did not affect the level of cell damage, and at 30°C and 37°C they increased it. The existence of temperature dependence of posthypertonic damage showed the involvement of the phospholipid component of the erythrocyte membrane in the process. Lower temperature is characterized by greater orderliness of lipids, its increase is accompanied by disorder and increased fluidity, and hence elasticity of the membrane. As a result, erythrocyte damage in posthypertonic shock is less at the temperature of 20–37°C. The addition of amphiphilic compounds at 0 and 10°C acts similarly to increasing the temperature, disorganizes the bilayer, increases the elasticity of the membrane and reduces damage during the transfer from hypertonic to isotonic solution. Above 20°C, the introduction of amphiphilic compounds leads not only to disorder, but also to the formation of mixed micelles consisting of phospholipids and amphiphilic molecules. This disrupts the bilayer, gives it instability and leads to increased damage of erythrocytes.

Key words: rabbit erythrocytes, amphiphilic compounds, posthypertonic shock, temperature

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1. Alvesa I, Stanevab G, Tessierac C, Salgadod GF, Nussac P. The Alves I, Staneva G, Tessier C, Salgado GF, Nuss P. The interaction of antipsychotic drugs with lipids and subsequent lipid reorganization investigated using biophysical methods. Biochim. Biophys. Acta Biomembr. 2011; 1808 (8): 2009–2018. DOI: 10.1016/j.bbamem.2011.02.021.
2. Bojic S, Murray A, Bentley BL, Spindler R, Pawlik P, Cordeiro JL, Bauer R, de Magalhães JP. Winter is coming: the future of cryopreservation. BMC Biol. 2021; 19 (1): 56. DOI: 10.1186/s12915-021-00976-8.
3. Chabanenko OO, Yershova NA, Orlova NV, Shpakova NM. Effect of sodium decyl sulfate and chlorpromazine on posthypertonic shock of mammalian red blood cells. Bìol. Tvarin. 2019; 21 (4): 84–90. DOI: 10.15407/animbiol21.04.084. (in Ukrainian)
4. Chabanenko O, Yershova N, Shpakova N. Adequacy of posthypertonic shock model to real cryopreservation conditions during deglycerolization of erythrocytes. Proc. 57^th ann. meet. “CRYO-2020”, 21–23 July 2020, USA. Cryobiol. 2020; 97: 276. DOI: 10.1016/j.cryobiol.2020.10.106.
5. Conrard L, Stommen A, Cloos AS, Steinkühler J, Dimova R, Pollet H, Tyteca D. Spatial relationship and functional relevance of three lipid domain populations at the erythrocyte surface. Cell. Physiol. Biochem. 2018; 51 (4): 1544–1565. DOI: 10.1159/000495645.
6. Durell SR, Ben-Naim A. Temperature dependence of hydrophobic and hydrophilic forces and interactions. J. Phys. Chem. 2021; 125 (48): 13137–13146. DOI: 10.1021/acs.jpcb.1c07802.
7. Ershova NA, Shpakova NM, Orlova NV, Ershov SS. Amphiphiles as tools for studying hypertonic cryohemolysis of mammalian erythrocytes. Bìol. Tvarin. 2014; 16 (2): 48–56. (in Ukrainian)
8. Färber N, Westerhausen C. Broad lipid phase transitions in mammalian cell membranes measured by Laurdan fluorescence spectroscopy. Biochim. Biophys. Acta Biomembr. 2022; 1864 (1): 183794. DOI: 10.1016/j.bbamem.2021.183794.
9. Habibi S, Lee HY, Moncada-Hernandez H, Gooding J, Minerick AR. Impacts of low concentration surfactant on red blood cell dielectrophoretic responses. Biomicrofluidics. 2019; 13 (5): 054101. DOI: 10.1063/1.5113735.
10. Jaferzadeh K, Sim M, Kim N, Moon I. Quantitative analysis of three-dimensional morphology and membrane dynamics of red blood cells during temperature elevation. Sci. Rep. 2019; 9: 14062. DOI: 10.1038/s41598-019-50640-z.
11. Klaiss-Luna MC, Manrique-Moreno M. Infrared spectroscopic study of multi-component lipid systems: A closer approximation to biological membrane fluidity. Membranes. 2022; 12 (5): 534. DOI: 10.3390/membranes12050534.
12. Muldrew K. The salting-in hypothesis of post-hypertonic lysis. Cryobiol. 2008; 57 (3): 251–256. DOI: 10.1016/j.cryobiol.2008.09.007.
13. Orlova NV, Shpakova NM. Mechanism of protective effect of amphiphilic compounds during hypertonic hemolysis of erythrocytes. Fiziol. Zh. 2006; 52 (5): 55–61. PMID: 17176840. (in Ukrainian)
14. Riske KA, Domingues CC, Casadei BR, Mattei B, Caritá AC, Lira RB, Preté PSC, de Paula E. Biophysical approaches in the study of biomembrane solubilization: quantitative assessment and the role of lateral inhomogeneity. Biophys. Rev. 2017; 9 (5): 649–667. DOI: 10.1007/s12551-017-0310-6.
15. Shpakova NM, Orlova NV. About the mechanism of mammalian erythrocytes osmotic stability. Probl. Cryobiol. Cryomed. 2020; 30 (4): 331–342. DOI: 10.15407/cryo30.04.331.
16. Steinkopf S, Schelderup AK, Gjerde HL, Pfeiffer J, Thoresen S, Gjerde AU, Holmsen H. The psychotropic drug olanzapine (Zyprexa^®) increases the area of acid glycerophospholipid monolayers. Biophys. Chem. 2008; 134 (1–2): 39–46. DOI: 10.1016/j.bpc.2008.01.003.
17. Wesołowska O, Michalak K, Hendrich AB. Direct visualization of phase separation induced by phenothiazine-type antipsychotic drugs in model lipid membranes. Mol. Membrane Biol. 2011; 28 (2): 103–114. DOI: 10.3109/09687688.2010.533706.