RESUMO
Glutathione (GSH) is an important cellular antioxidant and has a critical role in maintaining the balance of cellular redox. In this study, we investigated the GSH biosynthesis genes involved in the elevation of endogenous GSH levels using an irradiation system with an irradiation dose rate of 1.78 mGy/h, which was about 40,000 times less than the dose rates used in other studies. The results showed that GSH levels were significantly increased in the low-dose (0.02 and 0.2 Gy) irradiated group compared to those in the non-irradiated group, but enzymatic antioxidants such as superoxide dismutase and catalase were not induced at any doses tested. The elevation in GSH was accompanied by elevated expression of glutamate-cysteine ligase modifier subunit, but no changes were observed in the expression of glutamate-cysteine ligase catalytic subunit and thioredoxin in de novo GSH synthesis. In the case of genes involved in the GSH regeneration cycle, the expression of glutathione reductase was not changed after irradiation, whereas glutathione peroxidase was only increased in the 0.2 Gy irradiated group. Collectively, our results suggest that the de novo pathway, rather than the regeneration cycle, may be mainly switched on in response to stimulation with long-term low-dose radiation in the spleen.
Assuntos
Glutamato-Cisteína Ligase/biossíntese , Glutationa/biossíntese , Glutationa/efeitos da radiação , Baço/efeitos da radiação , Animais , Catalase/efeitos da radiação , Raios gama , Glutamato-Cisteína Ligase/genética , Glutamato-Cisteína Ligase/efeitos da radiação , Glutationa/metabolismo , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , RNA Mensageiro/biossíntese , Baço/metabolismo , Superóxido Dismutase/efeitos da radiação , TiorredoxinasRESUMO
OBJECTIVES: The relationships between alpha-tocopherol, pro-oxidant and antioxidant enzyme status, and radiation toxicity were studied in stage II, III, and IVA oral squamous cell carcinoma patients. The low levels of malondialdehyde and increased activities of antioxidant enzymes were correlated with decreased oxidative stress by alpha-tocopherol in oral cancer patients treated with radiotherapy. The objective of the present study was to evaluate the effect of alpha-tocopherol on oxidant-antioxidant enzyme status in oral squamous cell carcinoma patients treated with radiotherapy. MATERIALS AND METHODS: The study included three groups with histologically confirmed oral squamous cell carcinoma patients (untreated), and they were further divided into two groups, viz., one consisting of patients who underwent radiotherapy alone (radiotherapy was given at the dosage of 6000 cGy in five fractions per week for a period of 6 weeks); and the other group treated with radiotherapy plus alpha-tocopherol supplementation (alpha-tocopherol was supplemented at a dosage of 400 IU/day) for the entire period of radiotherapy. RESULTS: A significant decrease ( P < 0.001) in malondialdehyde levels and increase in activities of antioxidant enzymes ( P < 0.001) in hemolysate were noticed in patients treated with radiotherapy and simultaneously supplemented with alpha-tocopherol when compared to radiation-treated patients. CONCLUSION: It was seen that alpha-tocopherol played a role in protecting against the damage caused by irradiation in oral squamous cell carcinoma patients treated with radiotherapy, by enhancing the antioxidant enzyme status and reducing the pro-oxidant status.
Assuntos
Antioxidantes/administração & dosagem , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/radioterapia , Neoplasias Bucais/enzimologia , Neoplasias Bucais/radioterapia , alfa-Tocoferol/administração & dosagem , Antioxidantes/metabolismo , Antioxidantes/efeitos da radiação , Carcinoma de Células Escamosas/tratamento farmacológico , Catalase/sangue , Catalase/efeitos dos fármacos , Catalase/efeitos da radiação , Feminino , Glucosefosfato Desidrogenase/sangue , Glucosefosfato Desidrogenase/efeitos dos fármacos , Glucosefosfato Desidrogenase/efeitos da radiação , Glutationa Peroxidase/sangue , Glutationa Peroxidase/efeitos dos fármacos , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/sangue , Glutationa Redutase/efeitos dos fármacos , Glutationa Redutase/efeitos da radiação , Glutationa Transferase/sangue , Glutationa Transferase/efeitos dos fármacos , Glutationa Transferase/efeitos da radiação , Humanos , Masculino , Malondialdeído/sangue , Malondialdeído/efeitos da radiação , Pessoa de Meia-Idade , Neoplasias Bucais/tratamento farmacológico , Espécies Reativas de Oxigênio/sangue , Espécies Reativas de Oxigênio/efeitos da radiação , Superóxido Dismutase/sangue , Superóxido Dismutase/efeitos dos fármacos , Superóxido Dismutase/efeitos da radiaçãoRESUMO
To assess the role of antioxidant defense system on exposure to ultra-violet-B (UV-B) radiation, the activities of antioxidant enzymes superoxide dismutase (SOD), ascorbic acid peroxidase (APX), glutathione reductase (GR) and guaiacol peroxidase (GPX), as well as the level of antioxidants ascorbic acid (AA) and alpha-tocopherol were monitored in cucumber (Cucumis sativus L. var long green) cotyledons. UV-B enhanced the activity of antioxidant enzymes as well as AA content, but decreased the level of alpha-tocopherol. Significant increase was observed in the activities of SOD and GPX. Analysis of isoforms of antioxidant enzymes by native-PAGE and activity staining revealed three isoforms of GPX in unexposed dark-grown cotyledons (control), and their intensity was enhanced by UV-B exposure. In addition, four new isoforms of GPX were observed in cotyledons after UV-B exposure. Although no new isoforms were observed for the other antioxidant enzymes, the activities of their existing isoforms were enhanced by UV-B.
Assuntos
Antioxidantes/metabolismo , Cucumis sativus/enzimologia , Cucumis sativus/efeitos da radiação , Ascorbato Peroxidases , Cotilédone/enzimologia , Cotilédone/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Isoenzimas/metabolismo , Isoenzimas/efeitos da radiação , Peroxidase/metabolismo , Peroxidase/efeitos da radiação , Peroxidases/metabolismo , Peroxidases/efeitos da radiação , Superóxido Dismutase/metabolismo , Superóxido Dismutase/efeitos da radiação , Raios UltravioletaRESUMO
When bacterial cells are stressed by a change in the environment, they respond by changing the activity of enzymes at both the transcriptional and post-transcriptional levels. The UVA component (400-315 nm) of solar radiation reaching the Earth's surface is one of the most common stresses encountered by bacteria in their environment. Bacteria have evolved various antioxidant defense systems to increase survival when subjected to the deleterious effects of UVA irradiation. Recently, UVA-induced cytotoxicity and oxidative damage have been shown to be dependent on radiation intensity and dose distribution, not just total energy dose. We now report that when Escherichia coli is subjected to continuous sublethal, low-fluence UVA irradiation (7.4 W/m(2)) while growing to stationary phase, it responds by changing the activity levels of hydroperoxidases (HPI, HPII), glutathione reductase and manganese superoxide dismutase. This leads to an attenuation of the growth-delay response and an increase resistance to lethal UVA irradiation. When E. coli is given a UVA dose of 135 kJ/m(2) delivered at a fluence rate of 50 W/m(2), extensive protein oxidation occurs which may contribute to the inhibition of key cellular enzymes, leading to cellular dysfunction, DNA damage and eventually death. Changes in antioxidant enzymes induced by low-fluence UVA irradiation do not confer greater protection from protein oxidation after a challenge dose of UVA irradiation delivered at a fluence rate of 50 W/m(2).
Assuntos
Escherichia coli/efeitos da radiação , Oxirredução/efeitos da radiação , Raios Ultravioleta , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Catalase/metabolismo , Catalase/efeitos da radiação , Morte Celular/efeitos da radiação , Escherichia coli/enzimologia , Escherichia coli/crescimento & desenvolvimento , Proteínas de Escherichia coli/metabolismo , Proteínas de Escherichia coli/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Modelos Biológicos , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Superóxido Dismutase/efeitos da radiaçãoRESUMO
Time-resolved flavin fluorescence anisotropy studies on glutathione reductase (GR) have revealed a remarkable new phenomenon: wild-type GR displays a rapid process of fluorescence depolarization, that is absent in mutant enzymes lacking a nearby tyrosine residue that blocks the NADPH-binding cleft. Fluorescence lifetime data, however, have shown a more rigid active-site structure for wild-type GR than for the tyrosine mutants. These results suggest that the rapid depolarization in wild-type GR originates from an interaction with the flavin-shielding tyrosine, and not from restricted reorientational motion of the flavin. A novel mechanism of fluorescence depolarization is proposed that involves a transient charge-transfer complex between the tyrosine and the light-excited flavin, with a concomitant change in the direction of the emission dipole moment of the flavin. This interaction is likely to result from side-chain relaxation of the tyrosine in the minor fraction of enzyme molecules in which this residue is in an unsuitable position for immediate fluorescence quenching at the moment of excitation. Support for this mechanism is provided by binding studies with NADP+ and 2'P-5'ADP-ribose that can intercalate between the flavin and tyrosine and/or block the latter. Fluorescence depolarization analyses as a function of temperature and viscosity confirm the dynamic nature of the process. A comparison with fluorescence depolarization effects in a related flavoenzyme indicates that this mechanism of flavin fluorescence depolarization is more generally applicable.
Assuntos
Flavinas/química , Flavinas/efeitos da radiação , Glutationa Redutase/química , Glutationa Redutase/efeitos da radiação , Espectrometria de Fluorescência/métodos , Algoritmos , Ativação Enzimática , Flavinas/análise , Glutationa Redutase/análise , Luz , Conformação ProteicaRESUMO
RP-1, a herbal preparation of Podophyllum hexandrum has already been reported to provide protection against whole body lethal gamma irradiation (10 Gy). It has also been reported to render radioprotection to germ cells during spermatogenesis. Present study was undertaken to unravel the cellular and molecular mechanism of action of RP-1 on testicular system in strain 'A' mice. Various antioxidant parameters such as thiol content, glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST) enzyme activity, lipid peroxidation (LPO) and total protein levels were investigated. Thiol content was seen to increase significantly (p < 0.05) in both RP-1 alone and RP-1 pretreated irradiated groups over the irradiated groups at 8, 16 and 24 h. Irradiation (10 Gy) significantly decreased GPx, GST and GR activity in comparison to untreated control but RP-1 treatment before irradiation significantly (p < 0.05) countered radiation-induced decrease in the activity of these enzymes. Radiation-induced LPO was also found to be reduced at all time intervals by RP-1 treatment before irradiation. As compared to irradiated group the protein content in testicular tissue was increased in RP-1 pretreated irradiated group at 4 and 16 h significantly (p < 0.05). Comets revealed by single-cell gel electrophoresis were significantly longer (p < 0.001) in irradiated mice than in unirradiated control. RP-1 treatment before irradiation, however, rendered significant increase (p < 0.05) in comet length over the corresponding control and irradiated group initially at 4 h but at later time points, this was reduced significantly (p < 0.01) as compared to the irradiated group. RP-1 treatment alone rendered shorter comets at 8, 16 and 24 h than irradiated groups (p < 0.001). This study implies that RP-1 offers radioprotection at biochemical and cytogenetic level by protecting antioxidant enzymes, reducing LPO and increasing thiol content.
Assuntos
Extratos Vegetais/farmacologia , Podophyllum , Protetores contra Radiação/farmacologia , Espermatogênese/efeitos da radiação , Irradiação Corporal Total , Animais , Ensaio Cometa , Raios gama , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/efeitos da radiação , Glutationa Transferase/efeitos da radiação , Peroxidação de Lipídeos/efeitos da radiação , Masculino , Camundongos , Camundongos Endogâmicos A , Fitoterapia , Extratos Vegetais/uso terapêutico , Plantas Medicinais , Proteínas/análise , Proteínas/efeitos da radiação , Compostos de Sulfidrila/efeitos da radiação , Fatores de TempoRESUMO
BACKGROUND AND PURPOSE: Both infrared and low-power laser have been applied to improve circulation, wound repair, and pain control. Infrared and low-power laser therapies have the potential for stimulating enzyme activities which might contribute to increased glutathione (GSH) concentration and provide protection against oxidative damage. This study investigated cell viability, and GSH and its related enzyme activities in rat hepatocytes after irradiation. METHODS: Hepatocytes were isolated from 8-week-old male Sprague-Dawley rats and the cultures were divided into infrared, laser, and control groups. The cells were treated with infrared and low-power laser at a distance of 35 cm for 20 minutes. The cell morphology, lactate dehydrogenase (LDH) leakage, lipid peroxidation, GSH concentration, GSH peroxidase, GSH reductase (GRd), and GSH S-transferase activities were measured after irradiation. RESULTS: The morphology and LDH leakage of hepatocytes in the irradiation groups did not differ significantly from those of the control group. After infrared irradiation, a significant decrease in thiobarbituric acid-reactive substances and an increase in GSH concentration were found after 48 hours of incubation compared to the control group (p < 0.05). Furthermore, laser irradiation resulted in a significant increase in GRd activity after 48 hours of incubation compared to the control group (p < 0.05). A 48-hour incubation period produced greater GRd activity in all groups compared to a 24-hour period (p < 0.05). CONCLUSIONS: Irradiation did not damage rat hepatocytes in this study. Infrared was shown to stimulate GSH production, while laser irradiation increased GRd activity.
Assuntos
Sobrevivência Celular/efeitos da radiação , Glutationa/efeitos da radiação , Raios Infravermelhos , Fígado/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Análise de Variância , Animais , Células Cultivadas , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Glutationa Transferase/metabolismo , Glutationa Transferase/efeitos da radiação , L-Lactato Desidrogenase/metabolismo , L-Lactato Desidrogenase/efeitos da radiação , Peroxidação de Lipídeos , Fígado/citologia , Fígado/enzimologia , Ratos , Ratos Sprague-DawleyRESUMO
Las formulaciones disponibles actualmente para uso dermatológico, basadas en sustancias antioxidantes tales como vitaminas C y E, entre otras, abundan con promesas de revertir el envejecimiento cutáneo. En el presente trabajo se realiza una revisión de los sistemas antioxidantes cutáneos, de la relación entre envejecimiento y daño oxidativo, así como de la evidencia disponible en cuanto al tratamiento con antioxidantes. La intención de este artículo es que el dermatólogo comprenda las bases fisiológicas de acción de los antioxidantes, para poder juzgar su utilidad con una mirada crítica
Assuntos
Humanos , Animais , Ácido Ascórbico/uso terapêutico , Antioxidantes , Espécies Reativas de Oxigênio , Pele , Envelhecimento da Pele , Raios Ultravioleta , Vitamina E , Ácido Ascórbico/farmacologia , Ácido Ascórbico/fisiologia , Administração Tópica , Antioxidantes , beta Caroteno , Catalase , Ensaios Clínicos como Assunto , Cosméticos , Fenômenos Fisiológicos da Pele , Glutationa Peroxidase/efeitos da radiação , Glutationa Peroxidase/fisiologia , Glutationa Redutase/efeitos da radiação , Glutationa Redutase/fisiologia , Glutationa Transferase/efeitos da radiação , Glutationa Transferase/fisiologia , Interleucinas , Ozônio , Peroxidase , Pele , Luz Solar , Superóxido Dismutase/efeitos da radiação , Superóxido Dismutase/fisiologia , Ubiquinona , Vitamina ERESUMO
The Syrian hamster Harderian gland (HG), an organ present in the male two secretory cell types (type-I and type-II cells), is physiologically exposed to high oxidative stress because of high concentrations of porphyrins and their precursor, 5-aminolevulinic acid. Because of its juxtaorbital location, the HG is accessible to light, and subject to phototoxic effects of these substances. After having previously demonstrated circadian rhythms in antioxidant enzymes, porphyric enzymes and oxidative damage of proteins and lipids, as well as influences of melatonin on these parameters, we have now studied the effects of continuous light (LL), which suppresses melatonin secretion by the pineal gland. Measurements were performed in two different circadian phases, in order to detect the presence or absence of day/night differences. In LL, no differences between circadian phases of subjective day and subjective night were demonstrable for 5-aminolevulinate synthase, 5-aminolevulinate dehydratase, porphobilinogen deaminase, or superoxide dismutase; temporal differences in glutathione reductase and catalase were markedly diminished, whereas all these parameters showed marked day/night differences in the rats exposed to a light/dark cycle of 14:10. In LL, oxidative damage to lipids was minimally effected, while protein damage was enhanced. LL also caused a reduction in the percentage of type-II cells. Therefore, cell differentiation in the HG does not seem to be controlled only by the androgen, but, unexpectedly, also by melatonin.
Assuntos
Catalase/efeitos da radiação , Glutationa Redutase/efeitos da radiação , Glândula de Harder/fisiologia , Luz , Porfirinas/biossíntese , Superóxido Dismutase/efeitos da radiação , Animais , Ritmo Circadiano/fisiologia , Ritmo Circadiano/efeitos da radiação , Cricetinae , Heme/biossíntese , MasculinoRESUMO
Metals and ultraviolet (UV) radiation are two environmental stressors that can cause damage to plants. These two types of stressors often impact simultaneously on plants and both are known to promote reactive oxygen species (ROS) production. However, little information is available on the potential parallel stress responses elicited by metals and UV radiation. Using the aquatic plant Lemna gibba, we found that copper and simulated solar radiation (SSR, a light source containing photosynthetically active radiation (PAR) and UV radiation) induced similar responses in the plants. Both copper and SSR caused ROS formation. The ROS levels were higher when copper was combined with SSR than when applied with PAR. Higher concentrations of copper plus PAR caused toxicity as monitored by diminished growth and chlorophyll content. This toxicity was more pronounced when copper was combined with SSR. Because the generation of ROS was also higher when copper was combined with SSR, we attributed this enhanced toxicity to elevated levels of ROS. In comparison to PAR-grown plants, SSR treated plants exhibited elevated levels of superoxide dismutase (SOD) and glutathione reductase (GR). These enzyme levels were further elevated under both PAR and SSR when copper was added at concentrations that generated ROS. Interestingly, copper treatment in the absence of SSR (i.e. copper plus PAR) induced synthesis of the same flavonoids as those observed in SSR without copper. Finally, addition of either dimethyl thiourea or GSH (two common ROS scavengers) lowered in vivo ROS production, alleviated toxicity and diminished induction of GR as well as accumulation of UV absorbing compounds. Thus, the potential of ROS being a common signal for acclimation to stress by both copper and UV can be considered.
Assuntos
Araceae/crescimento & desenvolvimento , Cobre/farmacologia , Fotossíntese/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/fisiologia , Tioureia/análogos & derivados , Aciltransferases/efeitos dos fármacos , Aciltransferases/metabolismo , Aciltransferases/efeitos da radiação , Araceae/efeitos dos fármacos , Araceae/metabolismo , Araceae/efeitos da radiação , Clorofila/metabolismo , Clorofila/efeitos da radiação , Flavanonas/biossíntese , Flavanonas/efeitos da radiação , Sequestradores de Radicais Livres/farmacologia , Glutationa/farmacologia , Glutationa Redutase/efeitos dos fármacos , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Fotossíntese/efeitos dos fármacos , Fotossíntese/efeitos da radiação , Espécies Reativas de Oxigênio/efeitos da radiação , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/efeitos da radiação , Energia Solar , Superóxido Dismutase/efeitos dos fármacos , Superóxido Dismutase/metabolismo , Superóxido Dismutase/efeitos da radiação , Tioureia/farmacologia , Raios UltravioletaRESUMO
Stipa capillata (Poaceae) seeds were harvested from a control area (displaying a gamma dose rate of 0.23 micro Sv h(-1)) (C plants) and from two contaminated areas (5.4 and 25 micro Sv h(-1)) on the Semipalatinsk nuclear test site (SNTS) in Kazakhstan. The plants were grown for 124 d in a greenhouse under controlled conditions and exposed to three different treatments: (0) control; (E) external gamma irradiation delivered by a sealed 137Cs source with a dose rate of 66 micro Sv h(-1); (E+I) E treatment combined with internal beta irradiation due to contamination by 134Cs and 85Sr via root uptake from the soil. The root uptake led to a contamination of 100 Bq g(-1) for 85Sr and 5 Bq g(-1) for 134Cs (of plant dry weight) as measured at harvest. The activity of SOD, APX, GR, POD, CAT, G6PDH, and MDHAR enzymes was measured in leaves. Under (0) treatment, all enzymes showed similar activities, except POD, which had higher activity in plants originating from contaminated areas. Treatment (E) induced an enhancement of POD, CAT, GR, SOD, and G6PDH activities in plants originating from contaminated areas. Only control plants showed any stimulation of APX activity. Treatment (E+I) had no significant effect on APX, GR, CAT, and POD activities, but MDHAR activity was significantly reduced while SOD and G6PDH activities were significantly increased. The increase occurred in plants from all origins for SOD, with a greater magnitude as a function of their origin, and it occurred only in plants from the more contaminated populations for G6PDH. This suggests that exposure to a low dose rate of ionizing radiation for almost a half century in the original environment of Stipa has led to natural selection of the most adapted genotypes characterized by an efficient induction of anti-oxidant enzyme activities, especially SOD and G6PDH, involved in plant protection against reactive oxygen species.
Assuntos
Glucosefosfato Desidrogenase/metabolismo , Oxirredutases/metabolismo , Poaceae/efeitos da radiação , Ascorbato Peroxidases , Partículas beta , Catalase/metabolismo , Catalase/efeitos da radiação , Césio/administração & dosagem , Radioisótopos de Césio/administração & dosagem , Cloretos/administração & dosagem , Raios gama , Glucosefosfato Desidrogenase/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Cazaquistão , NADH NADPH Oxirredutases/metabolismo , NADH NADPH Oxirredutases/efeitos da radiação , Oxirredutases/efeitos da radiação , Peroxidases/metabolismo , Peroxidases/efeitos da radiação , Poaceae/enzimologia , Poaceae/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Poluentes Radioativos do Solo/administração & dosagem , Estrôncio/administração & dosagem , Radioisótopos de Estrôncio/administração & dosagem , Superóxido Dismutase/metabolismo , Superóxido Dismutase/efeitos da radiaçãoRESUMO
The protective role of superoxide dismutases (SODs) against ionizing radiation, which generates reactive oxygen species (ROS) harmful to cellular function, was investigated in the wild-type and in mutant yeast strains lacking cytosolic CuZnSOD (sod1Delta), mitochondrial MnSOD (sod2Delta), or both SODs (sod1Deltasod2Delta). Upon exposure to ionizing radiation, there was a distinct difference between these strains in regard to viability and the level of protein carbonyl content, which is the indicative marker of oxidative damage to protein, intracellular H2O2 level, as well as lipid peroxidation. When the oxidation of 2',7'-dichlorofluorescin was used to examine the hydroperoxide production in yeast cells, the SOD mutants showed a higher degree of increase in fluorescence upon exposure to ionizing radiation as compared to wild-type cells. These results indicated that mutants deleted for SOD genes were more sensitive to ionizing radiation than isogenic wild-type cells. Induction and inactivation of other antioxidant enzymes, such as catalase, glucose 6-phosphate dehydrogenase, and glutathione reductase, were observed after their exposure to ionizing radiation both in wild-type and in mutant cells. However, wild-type cells maintained significantly higher activities of antioxidant enzymes than did mutant cells. These results suggest that both CuZnSOD and MnSOD may play a central role in protecting cells against ionizing radiation through the removal of ROS, as well as in the protection of antioxidant enzymes.
Assuntos
Saccharomyces cerevisiae/efeitos da radiação , Superóxido Dismutase/efeitos da radiação , Catalase/metabolismo , Catalase/efeitos da radiação , Sistema Livre de Células , Ativação Enzimática/efeitos da radiação , Glucosefosfato Desidrogenase/metabolismo , Glucosefosfato Desidrogenase/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/análiseRESUMO
Methyl 2-nitroimidazole-1-acetohydroxamate (KIN-804) is a 2-nitroimidazole derivative containing a hydroxamate side chain designed to enhance the radiosensitization response of hypoxic cells. The possible sensitization of tumor tissue by KIN-804 can be evaluated through investigation of the levels of the free radical scavengers; namely, glutathione (GSH) and its complex enzyme system including glutathione reductase (GR) and glutathione peroxidase (GSH-Px), as well as glucose-6-phosphate dehydrogenase (G-6-PD). Female albino mice were inoculated with Ehrlich carcinoma in the thigh. Administration of KIN-804 (i.p. 80 mg/kg body weight) was carried out 20 min before localized irradiation of 10 Gy. The data revealed that KIN-804 administration, followed or not by gamma irradiation, resulted in a significant decrease in GSH content in tumor tissues associated with inhibition in GR and G-6-PD activities. Blood GSH-Px was enhanced in tumor inoculated mice and the administration of KIN-804 returned it to the normal value. These changes were more noticeable in tumor bearing mice exposed to both KIN-804 and irradiation.
Assuntos
Carcinoma de Ehrlich/metabolismo , Sequestradores de Radicais Livres/farmacologia , Ácidos Hidroxâmicos/farmacologia , Nitroimidazóis/farmacologia , Radiossensibilizantes/farmacologia , Animais , Feminino , Glucosefosfato Desidrogenase/metabolismo , Glucosefosfato Desidrogenase/efeitos da radiação , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Camundongos , OxirreduçãoRESUMO
We examined the elevation of glutathione (GSH) levels in the liver of C57BL/6 female mice after low-dose r-ray irradiation and its inhibitory effect on CClI4-induced liver damage. The liver GSH level increased soon after irradiation with 50 cGy of gamma-rays, reached a maximum at around 12 post-treatment, and returned almost to the control level by 24 h. The activities of glutathione reductase, and glutathione peroxidase also showed the same pattern of change, while the activity of gamma-glutamylcysteine synthetase showed a gradual increase up to 24 h. The effect of pre-irradiation on CCl4-induced liver damage was also investigated. The activities of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase in serum were markedly increased 12 h post-treatment with CCl4. Both increases were significantly suppressed by a single low-dose pre-irradiation. Malondialdehyde, a marker of lipidperoxidation, was also greatly elevated after CCl4 treatment, and its increase was suppressed by irradiation. These results suggest low-dose gamma-ray irradiation might be effective for the prevention of and/or therapy of various reactive oxygen species-related diseases including cancer.
Assuntos
Intoxicação por Tetracloreto de Carbono/metabolismo , Glutationa/metabolismo , Fígado/efeitos da radiação , Animais , Intoxicação por Tetracloreto de Carbono/patologia , Feminino , Raios gama , Glutamato-Cisteína Ligase/metabolismo , Glutamato-Cisteína Ligase/efeitos da radiação , Glutationa/efeitos da radiação , Glutationa Peroxidase/metabolismo , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/metabolismo , Glutationa Redutase/efeitos da radiação , Cinética , Fígado/metabolismo , Fígado/patologia , Camundongos , Camundongos Endogâmicos C57BL , Fatores de TempoAssuntos
Glutationa Redutase/antagonistas & inibidores , Hematoporfirinas/farmacologia , Indóis/farmacologia , Compostos Organometálicos/farmacologia , Radiossensibilizantes/farmacologia , Catalase/farmacologia , Eritrócitos/enzimologia , Glutationa Redutase/sangue , Glutationa Redutase/efeitos da radiação , Humanos , Cinética , NADP/farmacologia , Fotólise , Saccharomyces cerevisiae/enzimologia , Superóxido Dismutase/farmacologiaRESUMO
In previous studies we have found that a single acute dose of ultraviolet radiation to murine skin causes a large degree of destruction of enzymic and non-enzymic antioxidants immediately after irradiation. In the present study, we wished to elucidate the recovery of antioxidants after a single dose of ultraviolet (UV) radiation. We measured antioxidants and lipid hydroperoxides (as a marker of membrane damage) in murine epidermis and the dermis at 0, 3, 12, 24, 72 and 120 h after exposure to UV radiation (25 J/cm2, UVA+UVB). Lipid hydroperoxides showed the highest values immediately after UV exposure and returned to control values within 24 h in both epidermis and dermis. The activities of catalase, glutathione peroxidase and glutathione reductase showed the lowest activities immediately after UV exposure; superoxide dismutase activities reached a minimum at 3 h postexposure. The pattern of recovery was different for each enzyme and for epidermis and dermis. The activities of superoxide dismutase and catalase decreased remarkably and recovered slowly. Superoxide dismutase in the dermis recovered full activity by 120 h and in the epidermis by 12 h. Catalase activity in both epidermis and dermis had returned to only 50% of control activity at 120 h, although the epidermis showed a temporary increase (to 93%) at 24 h. Glutathione peroxidase and glutathione reductase were slightly decreased immediately after irradiation, recovered to 100% at 3 h and then increased to 200-250% in both the epidermis and the dermis at various times; values had returned to 100% in epidermis by 120 h but remained elevated in dermis.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Antioxidantes/efeitos da radiação , Epiderme/efeitos da radiação , Peróxidos Lipídicos/efeitos da radiação , Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Animais , Antioxidantes/análise , Ácido Ascórbico/análise , Ácido Ascórbico/efeitos da radiação , Catalase/análise , Catalase/efeitos da radiação , Ácido Desidroascórbico/análise , Ácido Desidroascórbico/efeitos da radiação , Epiderme/metabolismo , Feminino , Glutationa/análogos & derivados , Glutationa/análise , Glutationa/efeitos da radiação , Dissulfeto de Glutationa , Glutationa Peroxidase/análise , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/análise , Glutationa Redutase/efeitos da radiação , Peróxidos Lipídicos/análise , Camundongos , Camundongos Pelados , Doses de Radiação , Pele/metabolismo , Superóxido Dismutase/análise , Superóxido Dismutase/efeitos da radiação , Fatores de Tempo , Ubiquinona/análogos & derivados , Ubiquinona/análise , Ubiquinona/efeitos da radiação , Vitamina E/análise , Vitamina E/efeitos da radiaçãoRESUMO
Nitrosoureas inactivate cellular glutathione reductase. N,'N'1,3-bis(trans-4-hydroxycyclohexyl)-N'-nitrosoureas (BCyNU), a nitrosourea reported to selectively inhibit glutathione reductase (GR) activity, was examined to determine if it could be used as a means to inhibit cellular levels of this enzyme in radiobiology studies. Confirmation of drug-induced inhibition of GR activity was demonstrated using a cell-free model system employing purified GR. Cellular studies with Chinese hamster V79A03 showed that BCyNU decreased cellular glutathione content concomitant with an inhibition of specific GR activity. Under relatively nontoxic conditions, cellular exposure to BCyNU (25 microM, 0.25 h) either before or after radiation treatment, increased cellular radiosensitivity with the optimum time for drug addition being immediately following radiation. At a BCyNU dosage which produced less than or equal to 5% cell toxicity, a marked decrease in radioresistance was characterized as a reduction in both Dq (24 +/- 1.5%) and Do (8 +/- 0.5%) concomitant with a 25 +/- 2% decrease in cellular glutathione reductase (GR) activity. At cytotoxic drug dosages (25 microM, 1 h; cell survival 79 +/- 7%), a marked radiosensitization manifested by a 1.25 +/- .07-fold reduction in the Dq was observed concomitant with a 49 +/- 4% decrease in GR activity. Using cells enriched in different stages of the cell cycle, BCyNU caused cell-age dependent cytotoxicity with preferential killing of cells in the radioresistant late-S-phase, a likely explanation for its radiosensitizing capabilities at high drug dosages. Data obtained at nontoxic drug dosages suggest that GR-inactivation may be an important component of cellular response to free-radical induced damage.
Assuntos
Glutationa Redutase/antagonistas & inibidores , Compostos de Nitrosoureia/farmacologia , Animais , Morte Celular , Linhagem Celular , Cricetinae , Cricetulus , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/efeitos da radiação , Radicais Livres , Glutationa Redutase/efeitos dos fármacos , Glutationa Redutase/efeitos da radiação , Especificidade por Substrato/efeitos dos fármacos , Especificidade por Substrato/efeitos da radiaçãoRESUMO
Local and general reactions have been studied after subjecting a rabbit eye to dispersed helium-neon laser radiation, 8 x 10(-6) wt/cm-2, 10 min daily for 10 days. There were recorded increase of blood filling of the uveal tract of the eye, changes in the indices of systemic hemodynamics, systolic and diastolic pressure, stroke and minute volume of blood volume, activation of separate links of antioxidant system connected both with oxidation-reduction of thiol disulfide system and the functioning of antiperoxide enzymes. The activation of antioxidant system has a local character, the changes in the retina of the eye and pigmented epithelium is expressed stronger than in the peripheral blood. The results obtained speak about relationship between the stimulating effect of helium-neon laser radiation on the rabbit eye and activation of antioxidant system.
Assuntos
Olho/efeitos da radiação , Lasers/efeitos adversos , Animais , Antioxidantes , Catalase/efeitos da radiação , Glutationa Redutase/efeitos da radiação , Masculino , Epitélio Pigmentado Ocular/enzimologia , Epitélio Pigmentado Ocular/efeitos da radiação , Coelhos , Retina/enzimologia , Retina/efeitos da radiação , Superóxido Dismutase/efeitos da radiação , Úvea/irrigação sanguínea , Úvea/efeitos da radiaçãoRESUMO
Reactive oxygen species are considered to play an important role in cutaneous pathology. Enzymic and non-enzymic antioxidants can prevent oxidative damage but may be overcome by strong pro-oxidative stimuli. The acute effect of a single exposure to near ultraviolet (UVA)/visible radiation (greater than 320 nm) on various skin antioxidants was examined in hairless mice immediately after irradiation. Impairment of cutaneous catalase and glutathione reductase activity was observed. Superoxide dismutase and glutathione peroxidase were not significantly influenced. Inhibition of catalase may render skin more susceptible to the damaging effects of hydrogen peroxide and its reaction products such as the hydroxyl radical. Partially diminished glutathione reductase activity is not accompanied by a change in reduced/oxidized glutathione level immediately after irradiation. There was a tendential (not statistically significant) decrease in cutaneous tocopherol, ubiquinol + ubiquinone 9 and ascorbic acid levels, either indicating direct photodestruction or consumption by reaction products of photooxidative stress. This partial impairment of the cutaneous antioxidant defense system by near ultraviolet/visible light, showing that the most susceptible component in skin is catalase, suggests possible pharmacological interventions.
Assuntos
Catalase/efeitos da radiação , Glutationa Peroxidase/efeitos da radiação , Glutationa Redutase/efeitos da radiação , Luz , Pele/efeitos da radiação , Superóxido Dismutase/efeitos da radiação , Raios Ultravioleta , Animais , Catalase/metabolismo , Feminino , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Camundongos , Camundongos Pelados , Valores de Referência , Pele/efeitos dos fármacos , Pele/enzimologia , Superóxido Dismutase/metabolismoRESUMO
The activity of superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione: dehydroascorbate oxidoreductase is lower in the bone marrow than in the liver. The changes in the cellular antioxidative enzymatic system during leg bones fracture and X-ray irradiation are more pronounced in the rat bone marrow, as compared to the liver. The data obtained are in keeping with selective bone-marrow radiosensitivity and with the effect of exogenous superoxide dismutase and glutathione as radioprotectors.