Journal
BULLETIN OF EXPERIMENTAL BIOLOGY AND MEDICINE
Volume 149, Issue 1, Pages 78-81Publisher
SPRINGER
DOI: 10.1007/s10517-010-0880-9
Keywords
superoxide; NO synthase inhibitors; cytometry; HeLa; ECV-304
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Funding
- Russian Academy of Sciences
- Russian Foundation for Basic Research [05-04-49261-a]
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The dynamics of intracellular levels of superoxide and NO after cell treatment with NO synthase inhibitors were studied in human cells expressing various NOS isoforms: endotheliocytes and ECV-304 (eNOS) and carcinoma cells and HeLa-G63 (iNOS). Cytometric analysis of changes in the cell fluorescence intensity was carried out using superoxide and NO fluorescent indicators (dihydroethidene and DAF-2-DA, respectively). Intracellular levels of superoxide decreased in HeLa-G63 and ECV-304 cells after their incubation in medium with aminoguanidine, L-NAME, and D-NAME. Intracellular NO level decreased only in HeLa-G63 cells after incubation in medium with aminoguanidine and L-NAME, but not D-NAME. The level of NO returned to normal after 7-h culturing in inhibitor-free medium, while the level of superoxide increased and remained high throughout 3 generations. Incubation of cells with D-NAME did not increase the intracellular level of superoxide. Presumably, high prolonged generation of superoxide is a delayed result of inhibition of NO synthesis in HeLa-G63 cells.
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