RESUMO
We evaluated pyrogallol cytotoxicity using Escherichia coli strains that express mammalian catalase gene derived from catalase mutant mice (Cs(b)) and wild-type (Cs(a)), and pyrogallol mutagenicity by Ames test. Pyrogallol was more toxic to Cs(b) rather than to Cs(a) (p < 0.05), while catalase, superoxide dismutase and ascorbic acid decrease the toxic effect. Pyrogallol also showed mutagenic effect (mutagenic index = 3.8 for 10 micromol pyrogallol/plate) while ascorbic acid (19.4% reduction, p < 0.001) and naringin (35.1% reduction, p < 0.001) played a protective role against it. Pyrogallol cytotoxicity and mutagenicity seem to be attributable, at least in part, to reactive oxygen species formation. This study also suggests that newly established catalase mutant E. coli is probably useful in hazard identification of oxidative chemicals.
Assuntos
Catalase/genética , Escherichia coli/efeitos dos fármacos , Pirogalol/farmacologia , Salmonella typhimurium/efeitos dos fármacos , Animais , Escherichia coli/genética , Camundongos , Camundongos Mutantes , Testes de Mutagenicidade , Salmonella typhimurium/genéticaRESUMO
The present study aimed to assess whether a newly constructed, catalase-deficient Escherichia coli strain that express mammalian catalase gene could be used to identify oxidative stress-generating chemicals. We tested l-3,4-dihydroxyphenylalanine (l-DOPA), a well-known agent that induces reactive oxygen species. We found that l-DOPA exposure reduced the survival of catalase-mutant E. coli in a dose-dependent manner, especially in the strains with lower catalase activities, implying the usefulness of these strains in assessment of oxidative chemicals.
Assuntos
Proteínas de Bactérias/genética , Bioensaio/métodos , Catalase/genética , Di-Hidroxifenilalanina/toxicidade , Monitoramento Ambiental/instrumentação , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Relação Dose-Resposta a Droga , Monitoramento Ambiental/métodos , Escherichia coli/enzimologia , Levodopa/farmacologia , Mutação , Estresse Oxidativo , Espécies Reativas de OxigênioRESUMO
Lawsone is an active naphthoquinone derivative isolated from henna (Lawsonia inermis L.), a widely used hair dye. Previous study on the toxicity of lawsone remains unclear since the involvement of oxidative stress and the kind of ROS (reactive oxygen species) involved have not been fully resolved yet. This present study reports the cytotoxic effects of lawsone and henna. We carried out CAT assay (a zone of inhibition test of bacterial growth and colony-forming efficiency test of transformant Escherichia coli strains that express mammalian catalase gene derived from normal catalase mice (Cs(a)) and catalase-deficient mutant mice (Cs(b))), Ames mutagenicity assay and H(2)O(2) generation assay. Lawsone generated H(2)O(2) slightly in phosphate buffer system and was not mutagenic in Ames assay using TA 98, TA 100 and TA 102, both in the absence and presence of metabolic activation. Lawsone exposure inhibited the growth of both Cs(a) and Cs(b) strains in a dose-dependent manner. Mean zone diameter for Cs(a) was 9.75+/-0.96 mm and 12.75+/-1.5 mm for Cs(b). Natural henna leaves did not show toxic effects, whereas two out of four samples of marketed henna products were shown toxicity effects. Catalase abolished zone of inhibition (ZOI) of marketed henna products, eliminated ZOI of lawsone in a dose-dependent manner and low concentration of exogenous MnSOD and Cu/ZnSOD eliminated the toxicity. Histidine and DTPA, the metal chelator; BHA and low concentration of capsaicin, the inducer of NADH-quinone reductase, effectively protected Cs(a) and Cs(b) against lawsone in this study. We suggest that lawsone cytotoxicity is probably mediated, at least in part, by the release of O(2)(-), H(2)O(2) and OH(-).
Assuntos
Antioxidantes/farmacologia , Catalase/metabolismo , Citoproteção/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Tinturas para Cabelo/toxicidade , Lawsonia (Planta) , Naftoquinonas/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Animais , Antioxidantes/metabolismo , Ácido Ascórbico/farmacologia , Hidroxianisol Butilado/farmacologia , Capsaicina/farmacologia , Catalase/genética , Relação Dose-Resposta a Droga , Escherichia coli/enzimologia , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Tinturas para Cabelo/isolamento & purificação , Histidina/metabolismo , Peróxido de Hidrogênio/metabolismo , Radical Hidroxila/metabolismo , Lawsonia (Planta)/química , Camundongos , Testes de Sensibilidade Microbiana , Mutação , Naftoquinonas/isolamento & purificação , Estresse Oxidativo/efeitos dos fármacos , Ácido Pentético/farmacologia , Extratos Vegetais/toxicidade , Superóxido Dismutase/metabolismo , Superóxidos/metabolismoRESUMO
Hydroquinone is a benzene-derived metabolite. To clarify whether the reactive oxygen species (ROS) are involved in hydroquinone-induced cytotoxicity, we constructed transformants of Escherichia coli (E. coli) strains that express mammalian catalase gene derived from catalase mutant mice (Cs(b), Cs(c)) and the wild-type (Cs(a)) using a catalase-deficient E. coli UM255 as a recipient. Specific catalase activities of these tester strains were in order of Cs(a) > Cs(c) > Cs(b) > UM255, and their susceptibility to hydrogen peroxide (H2O2) showed UM255 > Cs(b) > Cs(c) > Cs(a). We found that hydroquinone exposure reduced the survival of catalase-deficient E. coli mutants in a dose-dependent manner significantly, especially in the strains with lower catalase activities. Hydroquinone toxicity was also confirmed using zone of inhibition test, in which UM255 was the most susceptible, showing the largest zone of growth inhibition, followed by Cs(b), Cs(c) and Cs(a). Furthermore, we found that hydroquinone-induced cell damage was inhibited by the pretreatment of catalase, ascorbic acid, dimethyl sulfoxide (DMSO), and ethylenediaminetetraacetic acid (EDTA), and augmented by superoxide dismutase (both CuZnSOD and MnSOD). The present results suggest that H2O2 is probably involved in hydroquinone-induced cytotoxicity in catalase-deficient E. coli mutants and catalase plays an important role in protection of the cells against hydroquinone toxicity.