Original ContributionPro-oxidative synergic bactericidal effect of NO: kinetics and inhibition by nitroxides
Graphical abstract
The combination of NO having bacteriostatic effect and H2O2 having moderate bactericidal activity results in a dramatic synergic toxicity toward microbial pathogens.
Section snippets
Chemicals
Diethylamine NONOate sodium salt (DEA/NO) was purchased from Cayman Chemicals (Ann Arbor, MI, USA); sodium nitroprusside (SNP), 2,2,6,6,-tetramethylpiperidine-N-oxyl (TPO), and 4-hydroxy-2,2,6,6,-tetramethylpiperidine-N-oxyl (TPL) were purchased from Sigma–Aldrich (St. Louis, MO, USA). 3-Carbamoyl-2,2,5,5-pyrrolidine-N-oxyl (3-CP) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). NO was purchased from Matheson Gas Products and was purified by passing the gas through a series of
The synergic effect of NO
Addition of NO to E. coli cells subjected to H2O2 induced a synergic cytocidal effect. Fig. 1 displays survival curves observed for exponentially growing E. coli at ca. 2×106 cells ml−1 subjected to H2O2 or NO-donor DEA/NO alone and in combination. NO alone induced a bacteriostatic [53], though not bactericidal, effect. Similarly, H2O2 at ca. 1 mM inhibited the growth rate but hardly caused cell killing. The growth arrest was documented also by respective termination of the increase in OD660 nm.
Discussion
Previous studies demonstrated opposite results and arrived at contradictory conclusions characterizing the effects of NO on microorganisms subjected to oxidative stress. Further elucidation of the NO effect is important because if it enhances the bactericidal effect of H2O2 it represents a therapeutic gain in combating invading pathogens by NO without harming host cells. Conversely, bacterial defense by NO from oxidative stress implicates the bacterial NO synthase in protection of such
Conclusions
This research demonstrates that (i) NO, even at sub-μM/min flux, exerts a dramatic pro-oxidative synergic effect, which greatly potentiates H2O2-induced killing of both gram-negative and gram-positive microbial cells; (ii) NO, in the absence of H2O2, induces a bacteriostatic effect in these cells; (iii) for A. viscosus and S. mutans, and A.a., the synergic effect of NO progressively increases with the duration of exposure, possibly with a concomitant progressive loss of cellular GSH; (iv) the
Acknowledgments
This work was supported by the Queens College Research Enhancement Funds and PSC-CUNY Research Awards. This work was completed using equipment in the Queens College Core Facility for Imaging, Cellular and Molecular Biology.
References (66)
- et al.
Sodium nitroprusside stimulates noradrenaline release from rat hippocampal slices in the presence of dithiothreitol
Brain Res.
(1996) - et al.
Dual effect of nitric oxide on the hyperpolarization-activated inward current (I-f) in sino-atrial node cells of the rabbit
J. Mol. Cell. Cardiol.
(1998) - et al.
Nitric oxide inhibits proliferation of human endothelial cells via a mechanism independent of cGMP
Atherosclerosis
(1999) - et al.
Down-regulation of survivin in nitric oxide-induced cell growth inhibition and apoptosis of the human lung carcinoma cells
J. Biol. Chem.
(2004) - et al.
The role of glutathione in nitric oxide donor toxicity to SN56 cholinergic neuron-like cells
Brain Res.
(2004) - et al.
Resolving the nitric oxide paradox in acute tissue damage
Gastroenterology
(1997) - et al.
Inhibition of inducible nitric oxide synthase ameliorates endotoxin-induced gut mucosal barrier dysfunction in rats
Gastroenterology
(1997) - et al.
The paradox of nitric oxide in cirrhosis and portal hypertension: too much, not enough
Hepatology
(2002) - et al.
Nitric oxide: cytotoxicity versus cytoprotection—how, why, when, and where?
Nitric Oxide
(1997) - et al.
Differential effects of nitric oxide synthase inhibitors on endotoxin-induced liver damage in rats
Gastroenterology
(1997)
Cellular antioxidant and pro-oxidant actions of nitric oxide
Free Radic. Biol. Med.
Nitric oxide-mediated cytoprotection of hepatocytes from glucose deprivation-induced cytotoxicity: involvement of heme oxygenase-1
Hepatology
Nitric oxide protects against alkyl peroxide-mediated cytotoxicity—further insights into the role nitric oxide plays in oxidative stress
Arch. Biochem. Biophys.
Nitric oxide (NO) protects against cellular damage by reactive oxygen species
Toxicol. Lett.
Nitric oxide donors modulate ferritin and protect endothelium from oxidative injury
Free Radic. Biol. Med.
Application of the nitric oxide donor SNAP to cardiomyocytes in culture provides protection against oxidative stress
Nitric Oxide
Nitric oxide and antimicrobial activity of reactive oxygen intermediates
Immunopharmacology
NONOates (1-substituted diazen-1-ium-1,2-diolates) as nitric oxide donors: convenient nitric oxide dosage forms
Methods Enzymol.
The chemistry and tumoricidal activity of nitric oxide hydrogen peroxide and the implications to cell resistance susceptibility
J. Biol. Chem.
NO-induced oxidative stress and glutathione metabolism in rodent and human cells
Free Radic. Biol. Med.
Kinetics of nitrosation of thiols by nitric-oxide in the presence of oxygen
J. Biol. Chem.
Kinetics of the reaction between nitric oxide and glutathione: implications for thiol depletion in cells
Free Radic. Biol. Med.
Roles of heme oxygenase-1 in the antiproliferative and antiapoptotic effects of nitric oxide on Jurkat T cells
Mol. Pharmacol.
Nitric oxide suppresses apoptosis in human colon cancer cells by scavenging mitochondrial superoxide anions
Int. J. Cancer
Role of calcium in nitric oxide-induced cytotoxicity: EGTA protects mouse oligodendrocytes
J. Neurosci. Res.
Nitric oxide at a low concentration protects murine macrophage RAW264 cells against nitric oxide-induced death via cGMP signaling pathway
Br. J. Pharmacol.
The role of nitric oxide in inflammation and immunity
Arthritis Rheum.
Opposite effects of pre- and postischemic treatments with nitric oxide donor on ischemia/reperfusion-induced renal injury
J. Pharmacol. Exp. Ther.
Nitric oxide has dual opposite roles during early and late phases of apoptosis in cerebellar granule neurons
Apoptosis
Opposite effects of nitric oxide donors on DNA single strand breakage and cytotoxicity caused by tert-butylhydroperoxide
Br. J. Pharmacol.
Nitric oxide and reactive oxygen species in vascular injury
Antioxidant mechanisms of nitric oxide against iron-catalyzed oxidative stress in cells
Antioxid. Redox Signaling
Nitric oxide co-operates with hydrogen peroxide in inducing DNA fragmentation and cell lysis in murine lymphoma cells
Biochem. J.
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These authors contributed equally to this work.