Continuous electrochemical monitoring of nitric oxide production in murine macrophage cell line RAW 264.7
Language English Country Germany Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Cell Line MeSH
- Time Factors MeSH
- Nitrites analysis chemistry metabolism MeSH
- Electrochemical Techniques instrumentation methods MeSH
- Lipopolysaccharides pharmacology MeSH
- Macrophages chemistry drug effects metabolism MeSH
- Mice MeSH
- Nitric Oxide analysis biosynthesis chemistry metabolism MeSH
- Nitric Oxide Synthase Type II metabolism MeSH
- Thiazines MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine MeSH Browser
- Nitrites MeSH
- Lipopolysaccharides MeSH
- Nitric Oxide MeSH
- Nitric Oxide Synthase Type II MeSH
- Thiazines MeSH
In this study, we realized the continual and long-term electrochemical detection of NO production by stimulated macrophages using modified porphyrinic microsensor. The NO release from RAW 264.7 cells stimulated by lipopolysaccharide started 5 h after the lipopolysaccharide administration. After reaching its maximum at the sixth hour, the stable level of NO production was observed between the seventh and 12th hour of the experiment. This phase was followed by a gradual decline in NO production. A close correlation between the NO signal detected with microelectrode and nitrite accumulation, which had been determined in supernatants removed from stimulated cells, was observed. This finding was utilized for the calibration of the electrochemical experiment. The presence of iNOS enzyme, which constitutes a main requirement for NO production by stimulated macrophages, was confirmed by Western blot analysis of iNOS protein expression at key time points of the corresponding electrochemical experiment. The capability of our microsensor to instantaneously monitor the changes in the NO production by stimulated RAW 264.7 cells was demonstrated by the immediate decrease in the signal due to NO as a response to the addition of iNOS inhibitor into the cell culture medium.
References provided by Crossref.org
Modulation of metabolic activity of phagocytes by antihistamines
