Enhanced Expression of iNOS in Human Endothelial Cells during Long-Term Culturing with Extracellular DNA Fragments

NO synthesis by endothelial cells plays an important role in normal function of the cardiovascular system. This work was designed to evaluate the role of variations in properties of extracellular DNA in the regulation of NO synthesis. We studied the effect of four DNA samples with various base seque...

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Bibliographic Details
Published in:Bulletin of experimental biology and medicine 2010-08, Vol.149 (2), p.191-195
Main Authors: Kostyuk, S. V, Smirnova, T. D, Efremova, L. V, Konkova, M. S, Alekseeva, A. Yu, Kameneva, L. V, Veiko, N. N
Format: Article
Language:eng ; rus
Subjects:
Biomedical and Life Sciences
Biomedicine
Cell Biology
Cell Culture Techniques
CG-DNA
DNA
DNA - metabolism
DNA Primers - genetics
Endothelial Cells - metabolism
Endothelium
eNOS
Extracellular Space - metabolism
Fluorescence
Gene expression
Gene Expression Regulation - physiology
Genes
Humans
iNOS
Internal Medicine
Laboratory Medicine
Metabolites
MyD88
Myeloid Differentiation Factor 88 - metabolism
Nitrates
Nitric oxide
Nitric Oxide - biosynthesis
Nitric Oxide Synthase Type II - metabolism
Pathology
Receptors, Cell Surface - metabolism
RNA
RNA, Messenger - metabolism
TLR9
Toll-Like Receptor 9 - metabolism
Umbilical Veins - cytology
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Summary:NO synthesis by endothelial cells plays an important role in normal function of the cardiovascular system. This work was designed to evaluate the role of variations in properties of extracellular DNA in the regulation of NO synthesis. We studied the effect of four DNA samples with various base sequences (50 ng/ml) on functional activity of endothelial cells HUVEC during 24-h culturing. Human DNA fragments with high content of CG repeats increased intracellular content of NO and its metabolites (nitrites and nitrates) and accelerated oxidation of nitrites to nitrates. Changes in the content of NO metabolites after 24-h culturing was shown to depend on the expression of gene for inducible, but not for endothelial NO synthase. Increased expression of inducible NO synthase positively correlated with an increase in the content of mRNA for the adapter protein MyD88, but did not depend on TLR9 gene expression that encodes protein receptor for CG-DNA recognition. The intracellular concentration of MyD88 mRNA did not depend on the content of TLR9 mRNA. The existence of a variety of DNA-binding receptors apart from TLR9 receptor on the surface of endothelial cells was hypothesized. Activation of these receptors by extracellular DNA fragments stimulates expression of the adapter protein MyD88.
ISSN:0007-4888
1573-8221
DOI:10.1007/s10517-010-0905-4