Biomedical Engineering Reference
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was mixed with 1 ml of Griess reagent. ). Incubated for 20 min and measured the absorption
of the chromophore at 540 nm on a spectrophotometer "Shimadzu" (Japan). NaNO
2
was used
as a standard for constructing the calibration curve. In addition, used to determine the test
system NO (R and D Systems) and Multiskan Antos 2020 at a wavelength of 540 nm. The
experimental results are expressed in relative units. The EPR spectra of samples of culture
medium, prepared as frozen columns with a diameter of 3 mm and a height of 30 mm,
recorded on a spectrometer "ESP-300" firm "Bruker-Analitishe-Messtechnik" (Germany), at
liquid nitrogen temperature. For the formation of NO followed by the appearance of the EPR
spectra signals of nitrosyl complexes of hemoglobin (Hb), which was added as a trap NO.
R
ESULTS AND
D
ISCUSSION
Monolayer cultures of ECs were treated with IFN - α at concentration 1х106 - 1х105
IU/ml. Results of testing of nitrite are presented on figure 1. It has appeared that after
treatment of ECs IFNα in concentration 1х105 IU/ml the amount of nitrite in cultural medium
has increased to 10-15 % by 24 o'clock in comparison with its amount in control not treated
IFN ECs. Treatment with IFNα in concentration 1х106 IU/ml resulted in to essential increase
in the level of nitrite in cultural medium in comparison with interferon used in concentration
1х105 IU/ml. And, it is necessary to notice that increase of level of nitrite was observed
already to 3 and 6 hours after using of IFN (approximately on 30 %), at the subsequent
cultivation of ECs there was a further increase in the level of nitrite which to 24 and 48 hours
exceeded control values on 50-60 %.
1,8
2
1,6
1,4
1,2
1
1,0
0,8
0
10
20
30
40
50
Time, hrs
Figure 1. Dynamics of nitric oxide production in human vascular endothelial cells after treatment with
interferon-α: 1 - concentration of IFN-α 1x10
5
IU / ml, 2 - concentration of IFN-α - 1x10
6
IU / ml.
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