Redox thiol status plays a central role in the mobilization and metabolism of nitric oxide in human red blood cells

We assessed the redox thiol status influence on nitric oxide (NO) metabolism and efflux in erythrocytes stimulated with acetylcholinesterase substrate (acetylcholine, ACh) and inhibitor (velnacrine maleate, VM). Erythrocyte suspensions from healthy donors were incubated with increasing concentration...

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Published in:Cell biology international 2009-03, Vol.33 (3), p.268-275
Main Authors: Lopes de Almeida, José Pedro, Carvalho, Filomena Almeida, Silva-Herdade, Ana Santos, Santos-Freitas, Teresa, Saldanha, Carlota
Format: Article
Language:English
Subjects:
Acetylcholine - pharmacology
Acetylcholinesterase
Acetylcholinesterase - metabolism
Cholinesterase Inhibitors - pharmacology
Dithiothreitol - pharmacology
Erythrocyte
Erythrocytes - drug effects
Erythrocytes - metabolism
Humans
Nitric oxide
Nitric Oxide - metabolism
Oxidation-Reduction
Peroxynitrous Acid - metabolism
Redox thiol status
S-Nitrosoglutathione - metabolism
S-nitrosothiols
Tacrine - analogs & derivatives
Tacrine - pharmacology
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Summary:We assessed the redox thiol status influence on nitric oxide (NO) metabolism and efflux in erythrocytes stimulated with acetylcholinesterase substrate (acetylcholine, ACh) and inhibitor (velnacrine maleate, VM). Erythrocyte suspensions from healthy donors were incubated with increasing concentrations of dithiothreitol (1–50 μM), in the presence and absence of acetylcholine/velnacrine (10 μM). Levels of NO, nitrite/nitrate, S-nitrosohemoglobin, peroxynitrite and S-nitrosoglutathione were determined by spectrofluorimetric and spectrophotometric methods. Dithiothreitol significantly mobilized NO toward nitrite/nitrate and S-nitrosoglutathione, and decreased the amount of NO efflux. Both ACh/VM induce changes on the levels of erythrocyte nitrite/nitrate dependent on the DTT concentration. Higher levels of peroxynitrite and S-nitrosoglutathione were seen with velnacrine in presence of DTT 1 and 50 μM. We concluded that dithiothreitol-induced activation of erythrocyte thiol status decreases NO efflux and allows greater intracellular NO mobilization onto different derivative molecules, both in the absence and presence of acetylcholinesterase substrate and inhibitor.
ISSN:1065-6995
1095-8355
DOI:10.1016/j.cellbi.2008.11.012