EVALUATION OF THE FORM AND DISTRIBUTION OF PHOSPHATIDYLSERINE IN HUMAN ERYTHROCYTES EXPOSED TO POSTHYPERTONIC SHOCK UNDER THE PROTECTION OF AMPHIPHILIC COMPOUNDS
O.E. Nipot, N.A. Yershova, O.O. Chabanenko, P.M. Zubov, N.M. Shpakova
Institute of Problems of Cryobiology and Cryomedicine, National Academy of Sciences of Ukraine, Kharkiv, Ukraine
DOI: https://doi.org/10.15407/fz70.02.059
Abstract
The search for protective substances that can be used during
red blood cell thawing and the study of their effects on red
blood cells contribute to increasing the number and quality of
viable cells after the cryopreservation cycle. We studied the
effect of posthypertonic shock and amphiphilic compounds on
the shape and eryptosis of human erythrocytes. The method of
flow cytometry was used, this allows analyzing two parameters
simultaneously, which increases the efficiency of research.
The shape was assessed by the sphericity index (SphI), and
eryptosis by the redistribution of phosphatidylserine to the
membrane outer surface. It has been shown that sodium
decylsulfate and chlorpromazine reduce erythrocyte damage
in posthypertonic shock by 3.6 and 4.2 times, respectively.
Sodium decylsulfate helps to preserve the shape of cells (SphI
coefficient remains the same), while when chlorpromazine is
used, the shape changes towards spherical (SphI coefficient
changes 2 times). The study of the level of Annexin V FITC
binding to phosphatidylserine in outer layer of membrane
revealed a concentration-dependent increase in fluorescence
when sodium decylsulfate was used, indicating a disorder of
the bilayer asymmetry. In contrast, chlorpromazine did not
change the distribution of phosphatidylserine. Comparison
of two parameters of cell viability - the sphericity coefficient
and anexin binding - allowed us to choose the conditions that
are optimal for the use of the studied protective substances.
Namely, it is advisable to use the lowest effective concentration
of sodium decylsulfate (200 mcmol/l) for protective purposes.
This ensures the preservation of the cell shape and minimal
impact on the membrane asymmetry.
Keywords:
erythrocyte; posthypertonic shock; amphiphilic compounds; sphericity coefficient; phosphatidylserine; flow cytometry.
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