Oxygen scavenging, NADH oxidase and metronidazole resistance in Helicobacter pylori.
Failure of triple-therapy regimes to eradicate Helicobacter pylori from the stomach is thought to be due to the occurrence of a metronidazole-resistant bacterial population. Exposure of metronidazole-resistant (MtzR) strains of H. pylori to an anaerobic environment causes the activation of metronidazole and the loss of resistance. Using metronidazole-sensitive (MtzS) clinical isolates, we selected mutants conferring resistance to metronidazole, which were used to investigate the effect of bacterial cell density upon the activation of metronidazole. The addition of metronidazole, at a final concentration of 10 mg/L, to MtzR cultures of a bacterial cell density >1 x 10(6) cfu/mL, caused a loss in viability. No loss in viability, however, occurred upon addition of metronidazole to MtzR cultures of a cell density of <1 x 10(6) cfu/mL. MtzS cultures lost viability irrespective of the initial cell density, indicating that oxygen scavenging at the site of metronidazole reduction may occur in these cultures. The ability of MtzS wild types, MtzR isogenic mutants and MtzR wild types to scavenge oxygen from the intracellular environment was investigated. H. pylori cultures contained NADH and NADPH oxidase activity. NADPH oxidase activity was always more than double the NADH oxidase activity. MtzR mutants possessed approximately one-third the NADH oxidase activity found in their respective MtzS parent wild types. MtzR wild types possessed a low NADH oxidase level similar to that found in the MtzR mutants. We propose that metronidazole resistance may be mediated through an inability of MtzR strains to remove oxygen from the site of metronidazole reduction, thereby preventing metronidazole activation.
Smith
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MA
Chemotherapy Research Unit, University of East London, Stratford Campus, UK.
Edwards
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DI
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002452:Cell Count; D004352:Drug Resistance, Microbial; D016480:Helicobacter pylori / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D008795:Metronidazole / Q000494:pharmacology*; D009097:Multienzyme Complexes / Q000378:metabolism* / Q000502:physiology; D009154:Mutation; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism* / Q000502:physiology; D019255:NADPH Oxidases / Q000378:metabolism; D012150:Polymorphism, Restriction Fragment Length
D000900:Anti-Bacterial Agents; D009097:Multienzyme Complexes; D008795:Metronidazole; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases