Activation of phagocytic cell NADPH oxidase by norfloxacin: a potential mechanism to explain its bactericidal action.
The mechanisms underlying the bactericidal power of fluoroquinolones against intracellular parasites in host macrophages remain poorly understood. We have analyzed the effect of norfloxacin, a fluoroquinolone antibiotic, on the production of reactive oxygen intermediates (O(2)(*-) and H(2)O(2)) and NADPH oxidase activity in mouse macrophages. The generation of anion superoxide (O(2)(*-)) was found to be significantly greater in macrophages incubated with norfloxacin than in untreated controls. This enhancing effect of norfloxacin was dose-dependent and reached maximal values within 10 min after its addition. The O(2)(*-) generated was mainly intracellular, as determined by the use of specific dyes, such as lucigenin and luminol, and able to diffuse freely through the cell membrane. Also, the production of H(2)O(2) was increased in macrophages in response to norfloxacin. The positive effect of norfloxacin was associated to an enhanced mobilization of NADPH oxidase subunits p47(phox) and p67(phox) from the cytosol to the plasma membrane in phagocytic cells. The effect of the antibiotic persisted in vivo for several hours. These data support the notion that norfloxacin inhibits mycobacterial growth within phagocytic cells by enhancing intracellular production of O(2)(*-) and other reactive oxygen species.
El Bekay
Rajaa
R
Departamento de Bioquímica Médica y Biología Molecular, Hospital Universitario Virgen Macarena, Universidad de Sevilla, Spain.
Alvarez
Moisés
M
Carballo
Modesto
M
Martín-Nieto
José
J
Monteseirín
Javier
J
Pintado
Elizabeth
E
Bedoya
Francisco J
FJ
Sobrino
Francisco
F
D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D002474:Cell-Free System; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D017737:Macrophages, Peritoneal / Q000201:enzymology* / Q000276:immunology; D051379:Mice; D019255:NADPH Oxidases / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology* / Q000276:immunology; D009643:Norfloxacin / Q000494:pharmacology*; D010587:Phagocytosis / Q000276:immunology; D017382:Reactive Oxygen Species / Q000276:immunology / Q000378:metabolism*
D000890:Anti-Infective Agents; D017382:Reactive Oxygen Species; D019255:NADPH Oxidases; D009643:Norfloxacin