[Erythropoietin prevents oxidative stress and lipid peroxidation process in human umbilical vein endothelial cells induced by tumor necrosis factor alpha].
According last research erythropoietin (Epo), glycoprotein known as a physiological regulator of erythropoiesis and used successfully to treat anemia, displays cytoprotective properties. It is supposed that the protective effect of EPO is based on its ability to oxidation-reduction stabilization of cells.


OBJECTIVE
The aim of the study was to examine the influence of oxidative stress and lipid peroxidation in human umbilical vein endothelial cells (HUVEC) induced by tumor necrosis factor-alpha (TNF-alpha).


METHODS
Human umbilical vein endothelial cells were cultured in a standard medium (M 199). The experiment was performed five times on the forth passage of HUVEC culture. For stimulation TNF-alpha was used in concentrations: 10, 20, 40 ng/ml (6 hours) and erythropoietin in concentrations: 10, 20, 40 IU/ml (24 hours). In the HUVEC lysate malonyldialdehyde (MDA), lipid hydroperoxides and reduction gluthation (GSH) concentrations were measured by HPLC method.


RESULTS
MDA concentration statistically significantly decreased from 16.48 +/- 1.21 to 14.40 +/- 0.72 nmol/mg protein and LOOH concentration from 73.00 +/- 5.44 do 68.86 +/- 1.89 nmol/mg protein in the cell's lysate due to the preincubation with Epo of HUVEC cells, stimulated with TNF-alpha. MDA and LOOH concentrations in the control culture were 12.91 +/- 1.02 nmol/mg protein and 57.80 +/- 6.16 nmol/mg protein respectively. The application of erythropoietin while the cells were being stimulated with TNF-alpha prevented the decrease in GSH concentration which was 34.77 +/- 0.70 nmol/mg protein in the control culture, 33.11 +/- 1.65 nmol/mg protein in the culture stimulated with TNF-alpha and 34.17 +/- 0.14 nmol/mg protein in culture preincubated with Epo and stimulated by TNF-alpha.


CONCLUSIONS
Erythropoietin (Epo) prevents oxidative stress and lipid peroxidation process in human umbilical vein endothelial cells (HUVEC) induced by tumor necrosis factor-alpha.
Szczepański
Marek
M
Department of Neonatology, Medical University of Bialystok, Poland. szczepanski5@gazeta.pl
Kamianowska
Monika
M
Wnuczko
Krzysztof
K
Skrzydlewska
Elzbieta
E
Wołczyński
Sławomir
S
D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D004921:Erythropoietin / Q000494:pharmacology* / Q000502:physiology; D005978:Glutathione / Q000187:drug effects; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects*; D008315:Malondialdehyde / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology* / Q000502:physiology; D014471:Umbilical Veins / Q000187:drug effects / Q000378:metabolism*
D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D004921:Erythropoietin; D008315:Malondialdehyde; D005978:Glutathione