424
N. A. Klenova and Z. P. Belousova
TABLE 1. Effect of Oleoyl Ethanolamine (IV) on Whole Donor Blood in the Presence and in the Absence of Lactic Acid
Experiment
Glucose absorption
Deformability, rel.
units
ATP, mM
DOXHb, %
OXHb, %
METHb, %
MBHb, %
(n = 15)
rate, mM/min
Control
10.16 ± 0.09
10.15 ± 0.12
16.07 ± 0.44
17.07 ± 0.29
51.80 ± 0.05
45.57 ± 0.05
2.12 ± 0.08
13.66 ± 0.16
0.76 ± 0.03
Compound IV
52.27 ± 0.02
P < 0.01
46.03 ± 0.05
P < 0.01
1.76 ± 0.07
P < 0.01
9.36 ± 0.39
P < 0.01
1.20 ± 0.06
P < 0.01
Lactic acid
5.71 ± 0.28
P < 0.01*
51.63 ± 0.03
P < 0.05*
45.33 ± 0.04
P < 0.01*
2.43 ± 0.08
P < 0.05*
14.01 ± 0.55
0.47 ± 0.03
P < 0.01*
Lactic acid + IV
10.02 ± 0.12
P < 0.01**
52.17 ± 0.04
P < 0.01**
45.81 ± 0.05
P < 0.01**
1.97 ± 0.03
P < 0.05**
9.91 ± 0.53
P < 0.01**
0.65 ± 0.03
P < 0.01**
Note: P* and P** indicate reliable differences from control and lactic acid test, respectively.
chloroform solution of compound IV was dried over calcium
chloride and evaporated on a rotor evaporator.
(Table 1). The mechanism of this destabilization is related to
an excess of protons and to a shift of the curve of hemoglo-
bin dissociation toward the formation of DOXHb, which is
accompanied by accelerated hemoglobin production and
penetration into the membrane, detrimentally affecting the
microviscosity characteristics.
The yield of compound IV in the form of a light-yellow
oil (crystallized on standing) was 6.5 g (65.6 %); Rf, 0.61; IR
spectrum (nmax, cm – 1): 3290 (O–H, alcohol), 3340 and 3150
(N–H, amide), 1650 (C=O, amide), 1530 (N–H, amide),
1400 (C – N, amide), 1350 (C=C, alkene), 1195, 1134, 1020
The introduction of oleoyl ethanolamine IV into the in-
cubation medium decreases the damaging effect of
lactoacidosis and reduces the rate of destabilization pro-
cesses in the presence of excess protons. There is a reliable
improvement of glucose transport, a decrease in the METHb
and MBHb fractions, and an increase in the cell
deformability.
1
(C=O, alcohol); H NMR spectrum (d, ppm): 3.905 (s, 1H,
OH), 0.93 (t, 3H, CH3), 1.19 – 1.27 (m, 7CH2), 1.92 – 2.2
(m, 2H), 1.16 – 1.57 (m, 5CH2), 2.14 – 2.20 (m, 2H),
5.35 – 5.40 (m, NH), 3.27 – 3.35 (2H), 3.47 – 3.58 (m, 2H),
5.20 – 5.28 (m, 2H).
EXPERIMENTAL BIOLOGICAL PART
Thus, compound IV favors protection of erythrocyte
cells under the condition of excess proton production. In ad-
dition, compound IV produces no damaging action on eryth-
rocytes by itself and, instead, stabilizes cell metabolism.
Human whole donor blood was incubated with oleoyl
ethanolamine (10 mg/ml) and lactic acid (7.5 mmole/ml) for
20 min at 37°C. The control sample was incubated with pure
solvents (ethyl alcohol and 0.154 M sodium chloride). In or-
der to maintain hematocrit, the volume of additives
amounted to 0.01 of the sample volume. The samples were
characterized by the rate of glucose absorption, ATP content,
hemoglobin state, and cell deformability [4 – 8].
REFERENCES
1. V. V. Bezuglov, M. Yu. Bobrov, and A. V. Archakov, Biokhimiya,
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(1998), pp. 34 – 37.
It was established that adding oleoyl ethanolamine (IV)
to the whole blood, while changing neither ATP content nor
the rate of glucose absorption by erythrocytes, reliably modi-
fies the ratio of hemoglobin forms: (i) increases the fraction
of deoxyhemoglobin (DOXHb) and oxyhemoglobin (OXHb)
at the expense of decreasing methemoglobin (METHb) frac-
tion; (ii) decreases the proportion of membrane-bound hemo-
globin (MBHb; and (iii) increases the cell deformability (Ta-
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(1996).
Lactoacidosis leads to accelerated destabilization of cells
during incubation, which is manifested by the decrease in the
rate of glucose absorption and deformability of the cells and
by the tendencyof the METHb and MBHb fractions to grow
7. Z. S. Toktamysova and N. Kh. Birzhanova, Biofizika, 35(6),
1019 – 1020 (1990).
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