RESUMO
Aquatic organisms are continuously exposed to environmental variations, which can lead to physiological and biochemical alterations. Leporinus macrocephalus, known as piavuçu, is a migratory species that may be exposed to variations in dissolved oxygen levels. Studies evaluating oxidative changes undergone by this species in these conditions are scarce. Therefore, this investigation aimed at evaluating oxidative alterations in L. macrocephalus exposed to different oxygen levels for 96 h: 6.12 ± 0.18, 3.99 ± 0.17, 3.22 ± 0.17, 2.47 ± 0.30 and 0.710 ± 0.07 mg L(-1). At the end of the experimental period, fish were euthanized and livers used to determine lipid hydroperoxides, thiobarbituric acid reactive substances, catalase, glutathione-S-transferase, superoxide dismutase and thiol groups, which are an indirect measure of reduced glutathione. Results indicated a decrease in the studied parameters in hypoxic situations, suggesting a possible metabolic depression.
Assuntos
Caraciformes/metabolismo , Oxigênio/metabolismo , Migração Animal , Animais , Antioxidantes/metabolismo , Catalase/metabolismo , Caraciformes/fisiologia , Doenças dos Peixes/metabolismo , Proteínas de Peixes/metabolismo , Glutationa/metabolismo , Glutationa Transferase/metabolismo , Hipóxia/metabolismo , Hipóxia/veterinária , Peroxidação de Lipídeos , Fígado/metabolismo , Oxirredução , Estresse Oxidativo , Oxigênio/administração & dosagem , Rios/química , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Thyroid hormones modulate haemoglobin and reactive oxygen species (ROS) production, leading to antioxidant changes. This study evaluated the antioxidant response to ROS in erythrocytes in hypothyroid and hyperthyroid rats. Wistar rats were divided into four groups: control; hyperthyroid (T4-12 mg 1(-1) in drinking water); sham operated (simulation of thyroidectomy); and hypothyroid (thyroidectomized). Four weeks after, blood was collected and haemoglobin and T(4) levels, lipid peroxidation (LPO), protein oxidation, superoxide dismutase (SOD), catalase (CAT) , glutathione S-transferase (GST) and glutathione peroxidase (GPx) activities, and total radical antioxidant potential (TRAP) were measured. SOD, CAT and GST immunocontent was evaluated. Haemoglobin levels were increased in hyperthyroid erythrocytes. LPO and carbonyls were augmented (65% and 55%, respectively) in hyperthyroid and reduced (31% and 56%, respectively) in hypothyroid group. SOD and CAT activities have not changed, as well as CAT immunocontent. TRAP was diminished in both hyperthyroid and hypothyroid groups (36% and 37%, respectively). GST activity and immunocontent, as well as GPx activity, were increased in hyper and hypothyroid rats. The data suggest that thyroid hormone changes determine ROS concentration changes and decrease of some antioxidant defences that would lead to a compensatory answer of the GST and GPx enzymes, which could be consider as credible biomarkers.
Assuntos
Antioxidantes/metabolismo , Eritrócitos/metabolismo , Hemoglobinas/metabolismo , Hormônios Tireóideos/metabolismo , Animais , Biomarcadores , Proteínas Sanguíneas/metabolismo , Catalase/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Hipertireoidismo/metabolismo , Hipotireoidismo/metabolismo , Peroxidação de Lipídeos , Masculino , Oxirredução , Estresse Oxidativo/fisiologia , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Tiroxina/sangueRESUMO
In 2015, glyphosate was classified as "Group 2A - probably carcinogenic to humans" by the International Agency for Research on Cancer (IARC). Therefore, public concerns about the environmental and health risks of this substance have rapidly increased. Considering its toxicokinetic characteristics, urinary levels of glyphosate could be a powerful tool for human biomonitoring. Nevertheless, the physicochemical properties of this molecule and the complexity of the matrix make this purpose particularly challenging. In order to solve this problem, the presented study describes a simple LC-MS/MS method for the quantification of glyphosate in human urine after pre-column derivatization with FMOC-Cl. Method development was focused on the optimization of the derivatization reaction in human urine, adjusting critical variables such as pH of borate buffer, FMOC-Cl concentration and derivatization time. Besides, chromatographic separation and spectrometric parameters were also established. The analytical method was fully validated according international guidelines for selectivity, carry over, linearity, accuracy, precision, lower limit of quantitation, matrix effect and stability under different conditions. All performance parameters were within the acceptance criteria. In addition, the method was successfully applied to 52 urine samples obtained from exposed subjects from northern Argentina, laying the foundation for future epidemiological studies.
Assuntos
Cromatografia Líquida/métodos , Fluorenos/química , Glicina/análogos & derivados , Espectrometria de Massas em Tandem/métodos , Adulto , Feminino , Glicina/química , Glicina/isolamento & purificação , Glicina/urina , Humanos , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Adulto Jovem , GlifosatoRESUMO
Phenylketonuria (PKU) is an autossomal recessive disease caused by phenylalanine-4-hydroxylase deficiency, which is a liver-specific enzyme that catalyzes the hydroxylation of l-phenylalanine (Phe) to l-tyrosine (Tyr). The deficiency of this enzyme leads to the accumulation of Phe in the tissues and plasma of patients. The clinical characterization of this disease is mental retardation and other neurological features. The mechanisms of brain damage are poorly understood. Oxidative stress is observed in some inborn errors of intermediary metabolism owing to the accumulation of toxic metabolites leading to excessive free radical production and may be a result of restricted diets on the antioxidant status. In the present study we evaluated various oxidative stress parameters, namely thiobarbituric acid-reactive species (TBA-RS) and total antioxidant reactivity (TAR) in the plasma of PKU patients. The activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were also measured in erythrocytes from these patients. It was observed that phenylketonuric patients present a significant increase of plasma TBA-RS measurement, indicating a stimulation of lipoperoxidation, as well as a decrease of plasma TAR, reflecting a deficient capacity to rapidly handle an increase of reactive species. The results also showed a decrease of erythrocyte GSH-Px activity. Therefore, it is presumed that oxidative stress is involved in the pathophysiology of the tissue damage found in PKU.
Assuntos
Estresse Oxidativo , Fenilcetonúrias/etiologia , Adolescente , Adulto , Criança , Pré-Escolar , Enzimas/sangue , Eritrócitos/enzimologia , Humanos , Peroxidação de Lipídeos , Fenilalanina/sangue , Fenilcetonúrias/sangue , Substâncias Reativas com Ácido Tiobarbitúrico/análiseRESUMO
Hyperthyroidism was induced in rats by l-thyroxine administration (12 mg/L in drinking water, 4 weeks). Animals were assessed hemodynamically, and heart, lung, and liver morphometry were performed. Lipid peroxidation (LPO) and protein oxidation (carbonyls) were measured in heart homogenates. It was quantified glutathione (GSH) metabolism, and antioxidant enzyme activities its and protein expression (by Western blot). At the end of treatment, it was observed cardiac hypertrophy, elevation of left ventricular systolic and end diastolic pressures, lung and liver congestion. LPO and carbonyls were increased in the hyperthyroid group, and GSH was decreased by 46% in the fourth week. Myocardial oxidative stress time course analysis revealed that it was increased in the second week of treatment. Antioxidant enzyme activities elevation was accompanied by protein expression induction in the hyperthyroid group in the fourth week. These results imply that hyperthyroidism generates myocardial dysfunction associated with oxidative stress inducing antioxidant enzyme activities and protein expression.
Assuntos
Antioxidantes/metabolismo , Glutationa/metabolismo , Hipertireoidismo/metabolismo , Miocárdio/enzimologia , Animais , Catalase/metabolismo , Glutationa Transferase/metabolismo , Cardiopatias/complicações , Hipertireoidismo/induzido quimicamente , Hipertireoidismo/complicações , Peroxidação de Lipídeos , Miocárdio/metabolismo , Oxirredução , Ratos , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , TiroxinaRESUMO
The present study was designed to evaluate the time course changes in peripheral markers of oxidative stress in a chronic HgCl2 intoxication model. Twenty male adult Wistar rats were treated subcutaneously daily for 30 days and divided into two groups of 10 animals each: Hg, which received HgCl2 (0.16 mg kg(-1) day(-1)), and control, receiving the same volume of saline solution. Blood was collected at the first, second and fourth weeks of Hg administration to evaluate lipid peroxidation (LPO), total radical trapping antioxidant potential (TRAP), and superoxide dismutase (Cu,Zn-SOD), glutathione peroxidase (GPx), glutathione-S-transferase (GST), and catalase (CAT). HgCl2 administration induced a rise (by 26%) in LPO compared to control (143 +/- 10 cps/mg hemoglobin) in the second week and no difference was found at the end of the treatment. At that time, GST and GPx were higher (14 and 24%, respectively) in the Hg group, and Cu,Zn-SOD was lower (54%) compared to control. At the end of the treatment, Cu,Zn-SOD and CAT were higher (43 and 10%, respectively) in the Hg group compared to control (4.6 +/- 0.3 U/mg protein; 37 +/- 0.9 pmol/mg protein, respectively). TRAP was lower (69%) in the first week compared to control (43.8 +/- 1.9 mM Trolox). These data provide evidence that HgCl2 administration is accompanied by systemic oxidative damage in the initial phase of the process, which leads to adaptive changes in the antioxidant reserve, thus decreasing the oxidative injury at the end of 30 days of HgCl2 administration. These results suggest that a preventive treatment with antioxidants would help to avoid oxidative damage in subjects with chronic intoxication.
Assuntos
Antioxidantes/análise , Eritrócitos/enzimologia , Peroxidação de Lipídeos/efeitos dos fármacos , Cloreto de Mercúrio/intoxicação , Estresse Oxidativo/efeitos dos fármacos , Peroxidases/sangue , Animais , Antioxidantes/metabolismo , Biomarcadores/sangue , Doença Crônica , Modelos Animais de Doenças , Luminescência , Masculino , Peroxidases/metabolismo , Ratos , Ratos Wistar , Fatores de TempoRESUMO
Cobalt chloride (CoCl2), a well-known inducer of heme oxygenase, produced a strong increase of in vivo rat liver chemiluminescence (QLV) 6 h after its administration. The activity of antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) was found to be significantly decreased 9 h after CoCl2 injection. Heme oxygenase activity increased 9 h after treatment, reaching a maximum value around 18 to 24 h after CoCl2 administration. This induction was preceded by a decrease in the intrahepatic GSH pool and an increase in hydrogen peroxide steady state concentration, both effects taking place several hours before induction of the heme-oxygenase. Co-administration of Sn-protoporphyrin IX, a potent inhibitor of heme oxygenase, completely prevented the enzyme induction, increasing the QLV levels. Administration of bilirubin, the end product of heme catabolism in mammals, prevented the heme oxygenase induction as well as the decrease in hepatic GSH and the increase of chemiluminescence when it was administered 2 h before CoCl2 treatment. These results support the proposal that the induction of heme oxygenase by cobalt chloride may be a general response to oxidant stress and, by increasing bilirubin levels, could constitute an important cellular defense mechanism against oxidative damage.
Assuntos
Bilirrubina/fisiologia , Heme Oxigenase (Desciclizante)/biossíntese , Fígado/enzimologia , Oxidantes/metabolismo , Animais , Cobalto/farmacologia , Indução Enzimática/efeitos dos fármacos , Feminino , Glutationa/análise , Peróxido de Hidrogênio/análise , Fígado/química , Medições Luminescentes , Ratos , Ratos WistarRESUMO
Fenofibrate, the hypolipidemic drug and peroxisome proliferator, was given to mice (0.23% w/w in the diet) during 1-3 weeks and enzyme activities, H2O2 concentration, and H2O2 production rate were determined. A maximal increase of 150% in liver/body weight ratio was observed after 3 weeks of treatment. Acyl-CoA oxidase, catalase and uricase activities were increased by 712%, 506% and 41% respectively by treatment with fenofibrate. Se- and non Se-glutathione peroxidase and Mn-superoxide dismutase activities were increased by 331%, 188% and 130% respectively in the liver of 2 weeks-treated mice. Cu-Zn superoxide dismutase activity was not affected by fenofibrate treatment. H2O2 steady-state concentration showed an increase of 89% after 2 weeks of treatment. H2O2 production rates, and the steady-state concentrations of the intermediates HO, R and ROO, calculated using experimental data, were higher in the liver of fenofibrate-treated mice than in control animals. According to our findings, the imbalance between H2O2 production and its degradation by its metabolizing enzymes during peroxisome proliferation, would result in an increased level of H2O2 steady-state concentration, with the resulting oxidative stress which may lead to the generation of oxidative damage and to the induction of liver carcinogenesis.
Assuntos
Fenofibrato/farmacologia , Peróxido de Hidrogênio/metabolismo , Hipolipemiantes/farmacologia , Fígado/metabolismo , Microcorpos/metabolismo , Acil-CoA Oxidase , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Peso Corporal , Catalase/metabolismo , Dieta , Feminino , Fenofibrato/administração & dosagem , Glutationa Peroxidase/metabolismo , Cinética , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Camundongos , Microcorpos/efeitos dos fármacos , Oxirredutases/metabolismo , Superóxido Dismutase/metabolismo , Urato Oxidase/metabolismoRESUMO
Fenofibrate, the hypolipidemic drug and peroxisome proliferator, was given to mice (0.23% w/w in the diet) during 1-3 weeks and H2O2 and TBARS steady state concentrations, liver chemiluminescence and antioxidant levels were measured. Administration of fenofibrate during 2 weeks induced an increase of 89% in H2O2 steady state concentration. Spontaneous chemiluminescence was decreased by 57% during fenofibrate treatment, while no significant effect was observed on TBARS concentration. Hydroperoxide-initiated chemiluminescence was decreased by 56% after 15 days of fenofibrate treatment, probably due to an increase in endogenous antioxidant levels. Total and oxidized glutathione increased gradually after fenofibrate administration, obtaining maximal increases of 67% and 58% respectively, after 22 days of treatment. An increase of 55% was found in ubiquinol levels in treated mice, as compared with the controls. alpha-tocopherol content was decreased by 51% in the liver of fenofibrate-treated mice. According to our findings, the high rate of H2O2 production associated with peroxisome proliferation, would not lead to an increase in lipid peroxidation. This can be explained by the presence of high levels of ubiquinols, which act as an antioxidant. The increased production of H2O2, would lead to DNA damage directly, and not through lipid peroxidation processes.
Assuntos
Antioxidantes/análise , Fenofibrato/administração & dosagem , Hipolipemiantes/administração & dosagem , Fígado/efeitos dos fármacos , Microcorpos/efeitos dos fármacos , Animais , Dano ao DNA , Feminino , Glutationa/análise , Peróxido de Hidrogênio/análise , Peroxidação de Lipídeos , Fígado/metabolismo , Fígado/ultraestrutura , Medições Luminescentes , Camundongos , Microcorpos/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/análiseRESUMO
X-linked adrenoleukodystrophy (X-ALD) is a hereditary disorder of peroxisomal metabolism biochemically characterized by the accumulation of very long chain fatty acids (VLCFA), particularly hexacosanoic acid (C26:0) and tetracosanoic acid (C24:0) in different tissues and in biological fluids. The disease is clinically characterized by central and peripheral demyelination and adrenal insufficiency, which is closely related to the increased concentrations of these fatty acids. However, the mechanisms underlying the brain damage in X-ALD are poorly known. Considering that free radical generation is involved in various neurodegenerative disorders, like Parkinson disease, multiple sclerosis and Alzheimer's disease, in the present study we evaluated various oxidative stress parameters, namely chemiluminescence, thiobarbituric acid reactive species (TBA-RS), total radical-trapping antioxidant potential (TRAP), and total antioxidant reactivity (TAR) in plasma of X-ALD patients, as well as the activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) in erythrocytes and fibroblasts from these patients. It was verified a significant increase of plasma chemiluminescence and TBA-RS, reflecting induction of lipid peroxidation, as well as a decrease of plasma TAR, indicating a deficient capacity to rapidly handle an increase of reactive species. We also observed a significant increase of erythrocytes GPx activity and of catalase and SOD activities in fibroblasts from the patients studied. It is therefore proposed that oxidative stress may be involved in pathophysiology of X-ALD.
Assuntos
Adrenoleucodistrofia/fisiopatologia , Estresse Oxidativo/fisiologia , Adrenoleucodistrofia/sangue , Adulto , Antioxidantes/metabolismo , Catalase/sangue , Células Cultivadas , Criança , Eritrócitos/enzimologia , Eritrócitos/metabolismo , Radicais Livres/metabolismo , Glutationa Peroxidase/sangue , Humanos , Superóxido Dismutase/sangue , Substâncias Reativas com Ácido Tiobarbitúrico/análiseRESUMO
Hydroperoxide-initiated chemiluminescence was standardized as a microassay to evaluate the occurrence of oxidative stress in human biopsies. Samples of 10 to 50 mg of rat liver or heart were homogenized, diluted in reaction medium, added with tert-butyl hydroperoxide, and assayed for chemiluminescence in a liquid scintillation counter in the out-of-coincidence mode. Optimal conditions for the assay were: 0.3 to 1.2 mg/mL of homogenate protein in 120 mM KCl, 30 mM phosphate buffer (pH 7.4), and 3 mM tert-butyl hydroperoxide at 30 degrees C. In these conditions, maximal chemiluminescence values were 550 +/- 30 and 1100 +/- 40 cps/mg protein, for liver and heart homogenates, respectively. Liver and heart homogenates were subjected to in vitro oxidative stresses such as supplementation with organic hydroperoxide or with enzymatic systems generating superoxide anion or hydrogen peroxide. Chemiluminescence was higher in the poststress samples than in the control ones. The ratio: poststress chemiluminescence/control chemiluminescence (B/A) was about 1.4 or higher for both tissues. Human heart biopsies were utilized to investigate the occurrence of oxidative stress after clinical situations associated to ischemia-reperfusion. B/A ratios were 2.1 +/- 0.4, 1.4 +/- 0.1, and 2.8 +/- 0.4 for human heart, liver, and skeletal muscle, respectively.
Assuntos
Fígado/metabolismo , Medições Luminescentes , Músculos/metabolismo , Miocárdio/metabolismo , Peróxidos/farmacologia , Animais , Biópsia , Radicais Livres , Humanos , Peróxido de Hidrogênio/metabolismo , Cinética , Fígado/efeitos dos fármacos , Masculino , Músculos/efeitos dos fármacos , Oxirredução , Ratos , Ratos Endogâmicos , Reperfusão , Superóxidos/metabolismo , terc-Butil HidroperóxidoRESUMO
Doxorubicin (1.2 mg/kg body weight) or 4'-epidoxorubicin (1.7 mg/kg body weight) was injected intravenously to rabbits twice a week during 7 to 8 weeks. Total doses were 17.9 +/- 0.2 mg and 24.4 +/- 0.3 mg, respectively. Heart, liver, muscle, and brain homogenates from treated and control animals were supplemented with 3 mM tert-butyl hydroperoxide and hydroperoxide-initiated chemiluminescence was measured. Heart homogenates from doxorubicin-treated rabbits showed an increased hydroperoxide-initiated chemiluminescence (77.2 +/- 3.9; expressed as cpm/mg protein X 10(-3]; whereas 4'-epidoxorubicin-treated rabbits did not exhibit changes (40.7 +/- 4.6) when both were compared with the untreated animals (41.3 +/- 3.0). Liver, muscle, and brain homogenates from doxorubicin and 4'-epidoxorubicin-treated animals showed a hydroperoxide-initiated chemiluminescence that was similar to the one from control animals. Microscopically, the total extent of the myocardial damage (as percentage of damaged myocytes) was markedly higher in the doxorubicin-treated rabbits (63.0 +/- 8.6) than in the 4'-epidoxorubicin-treated group (34.6 +/- 5.0); being both values higher than the one corresponding to control animals (8.0 +/- 1.1). The subendocardial areas of the septum and of the left ventricle were highly sensitive to doxorubicin damage. Hydroperoxide-initiated chemiluminescence of whole heart homogenate correlated statistically with the microscopic tissue damage in the subendocardial and intramural areas of the right ventricle. It is concluded that chronic administration of doxorubicins lead to oxidative stress of the myocardium and that 4'-epidoxorubicin produces less severe oxidative stress and less extensive myocardial damage than those provoked by lower doses of doxorubicin.
Assuntos
Doxorrubicina/toxicidade , Coração/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Animais , Epirubicina/farmacologia , Feminino , Radicais Livres , Medições Luminescentes , Masculino , Especificidade de Órgãos , Oxirredução , CoelhosRESUMO
Acetaminophen was given to mice at a single dose of 375 mg/kg. In situ liver chemiluminescence, H2O2 steady-state concentration, and the liver concentrations of total and oxidized glutathione were measured 15, 30, and 60 min after acetaminophen administration. Increases of 145% and 72% in spontaneous chemiluminescence and H2O2 concentration were observed 15 min after the injection, respectively. Total glutathione was decreased by acetaminophen administration at all the times studied. The maximal decrease, 83%, was found 60 min postinjection. The ratio GSH/GSSG was found significantly decreased at all the times studied. Microsomal superoxide production was increased by 2.4-fold by addition of acetaminophen. The activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase were determined. Catalase was slightly inhibited (30%) 15 min after acetaminophen administration. No significant changes were found in superoxide dismutase activity. Se and non-Se glutathione peroxidase activities were decreased by 40% and 53% respectively, 15 min after acetaminophen administration. The decrease in catalase and glutathione peroxidase would result in an increased steady state level of H2O2 and hydroperoxides, contributing to cell injury. Damaged hepatocytes were observed, and severe lesions and necrosis appeared 60 min after acetaminophen administration. Our results indicate the occurrence of oxidative stress as a possible mechanism for acetaminophen-induced hepatotoxicity.
Assuntos
Acetaminofen/toxicidade , Fígado/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/toxicidade , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Relação Dose-Resposta a Droga , Feminino , Glutationa/análogos & derivados , Glutationa/metabolismo , Dissulfeto de Glutationa , Peróxido de Hidrogênio/metabolismo , Cinética , L-Lactato Desidrogenase/sangue , Fígado/efeitos dos fármacos , Fígado/patologia , Medições Luminescentes , Camundongos , Superóxidos/metabolismoRESUMO
Sepsis, as infection associated to systemic manifestations, was produced in rats by cecal ligation and double perforation. Sham-operated rats were used as controls. The spontaneous chemiluminescence of rat adductor muscle and liver were measured at 6, 12, 24, and 30 h after the surgical procedure. Muscle chemiluminescence showed a maximal increase of about twofold (control emission 10 +/- 1 cps/cm2) after 6-12 h of sepsis, while liver chemiluminescence increased by about 80% (control emission: 11 +/- 1 cps/cm2) after 24 h of sepsis. The activities of muscle antioxidant enzymes were found maximally diminished after 12 h of sepsis: 46% decrease for Mn-superoxide dismutase, 83% decrease for catalase, and 55% decrease for glutathione peroxidase. In liver, only catalase activity showed a 52% decrease after 24 h of sepsis. State 3 oxygen uptake of muscle mitochondria with either malate-glutamate or succinate as substrates was 40% decreased after 12 h of sepsis in both cases. State 4 oxygen uptake of muscle mitochondria was not affected. The rate of H2O2 production of muscle mitochondria after 12 h of sepsis with either malate-glutamate or succinate as substrates was increased about 2.5 times but was not affected when assayed in the presence of as rotenone and antimycin. The oxygen uptake of liver mitochondria isolated from septic rats did not show differences as compared with those of control rats after 6 to 24 h of sepsis. Oxidative stress appears to occur in skeletal muscle early at the onset of the septic syndrome, with inhibition of active mitochondrial respiration and inactivation of antioxidant enzymes.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Peroxidação de Lipídeos , Fígado/metabolismo , Músculos/metabolismo , Sepse/metabolismo , Análise de Variância , Animais , Catalase/metabolismo , Ceco/cirurgia , Feminino , Glutationa Peroxidase/metabolismo , Peróxido de Hidrogênio/metabolismo , Cinética , Medições Luminescentes , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias Musculares/metabolismo , Consumo de Oxigênio , Ratos , Ratos Sprague-Dawley , Valores de Referência , Superóxido Dismutase/metabolismo , Fatores de TempoRESUMO
Fourteen randomized patients undergoing myocardial revascularization were divided into group A standard hypothermic cardioplegic solution) and group B (the same cardioplegic solution supplemented with deferoxamine 1000 mg/L). In all patients myocardial biopsy specimens were obtained before ischemia and during reperfusion and were assessed for chemiluminescence (to indirectly determine oxygen-free radical activity) and for electron microscopic studies. Chemiluminescence in group A showed a photoemission of 36.5 +/- 1.5 cpm/mg protein X10(-3) for the preischemia samples and 72 +/- 5.7 cpm/mg protein X10(-3) for the reperfusion samples (p less than 0.01). In the patients who received deferoxime (group B), values for chemiluminescence for preischemia and reperfusion samples were not significantly different. Electron microscopic studies showed a significant increase in grade 4 (severely damaged) mitochondria in reperfusion biopsy specimens from both groups as compared with preischemia samples. However, reperfusion samples from group B showed a better preservation of myocardial cells with marked reduction of grade 4 (severely damaged) mitochondria. These results support the hypothesis that oxygen-free radicals are responsible in part for the production of reperfusion injury in the human heart. They suggest that this mechanism may be at least partially controlled by adding an iron chelating agent such as deferoxime.
Assuntos
Soluções Cardioplégicas , Desferroxamina/administração & dosagem , Coração/efeitos dos fármacos , Reperfusão Miocárdica , Ponte de Artéria Coronária , Humanos , Peróxido de Hidrogênio/farmacologia , Medições Luminescentes , Pessoa de Meia-Idade , Mitocôndrias Cardíacas/ultraestrutura , Miocárdio/metabolismo , Miocárdio/ultraestrutura , Volume SistólicoRESUMO
Forty consecutive patients undergoing myocardial revascularization were divided into two equal groups: group 1 received standard cardioplegic solution, and group 2 received a solution containing mannitol, 59.8 mmol/L. In 6 patients in each group, myocardial biopsies were done before ischemia and at the time of reperfusion. Samples were assessed by chemiluminescence to determine oxidative stress and by electron microscopic studies. A significant reduction in atrial arrhythmias was observed in the mannitol group. Chemiluminescence in group 1 showed a photoemission of 37.6 +/- 3.5 cpm/mg of protein x 10(-3) for the preischemia samples and 74.8 +/- 16 cpm/mg of protein x 10(-3) for the reperfusion samples (p less than 0.001). In group 2, the values for chemiluminescence were 37.7 +/- 3.4 cpm/mg of protein x 10(-3) and 40 +/- 6.1 cpm/mg of protein x 10(-3), respectively (p = not significant). Electron microscopic studies showed, for group 1, increased grades of damaged mitochondria in the reperfusion biopsy specimens compared with the preischemia biopsy specimens (p less than 0.01). In group 2, differences for damaged mitochondria were not significant. These results support the hypothesis that mannitol reperfusate significantly reduces myocardial damage in patients undergoing open heart procedures. They also suggest that this protective effect may be in part secondary to the antioxidant property of mannitol, although other mechanisms may have accounted for or contributed to the improved outcome after ischemia.
Assuntos
Soluções Cardioplégicas/uso terapêutico , Parada Cardíaca Induzida , Manitol/uso terapêutico , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Biópsia , Radicais Livres , Humanos , Medições Luminescentes , Microscopia Eletrônica , Pessoa de Meia-Idade , Miocárdio/patologia , Estudos Prospectivos , Distribuição AleatóriaRESUMO
The heme precursor 5-aminolevulinic acid (ALA), acting as a prooxidant, has been proposed to underlie the clinical manifestations of various porphyric disorders. Accordingly, ALA-generated oxyradicals where shown to cause oxidative lesions in biomolecules and isolated cell organelles and to release iron from ferritin. In rats, administered ALA triggered oxidative stress in liver, brain and red muscles. We now study the correlation between the plasma antioxidant capacity and tissue oxidative damage, after acute (one and two doses) and prolonged (eight doses) ALA treatment of rats (one dose of ALA = 40 mg/kg body weight). The in situ spontaneous chemiluminescence intensity increased 5-fold in brain, 50% in liver and 4-fold in soleus muscle upon two dose-treatment, indicating tissue response to oxidative injury by ALA. Chemiluminescence reached the highest intensity after one or two doses of ALA and decreased after eight doses in all tissues. The plasma trapping capacity, evaluated by the luminol/2-amidinopropane system, gave a parallel response: maximum values after two doses and decreased values after prolonged treatment. After eight doses, the ALA concentration was found to be 3-fold above the normal value in plasma, 48% higher in liver and 38% higher in total brain. These data indicate that the plasma antioxidant system responds to ALA treatment and is correlated with tissue chemiluminescence. In vitro studies showed that ALA does not interfere with the antioxidant plasma capacity, neither promoting oxidation of plasma elements nor binding to plasma proteins.
Assuntos
Ácido Aminolevulínico/farmacologia , Antioxidantes/metabolismo , Estresse Oxidativo , Amidinas/química , Amidinas/metabolismo , Ácido Aminolevulínico/análise , Ácido Aminolevulínico/sangue , Animais , Ácido Ascórbico/química , Ácido Ascórbico/metabolismo , Proteínas Sanguíneas/efeitos dos fármacos , Proteínas Sanguíneas/metabolismo , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Cromanos/química , Cromanos/metabolismo , Glutationa/química , Glutationa/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Medições Luminescentes , Luminol/química , Luminol/metabolismo , Masculino , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Oxirredução , Plasma/efeitos dos fármacos , Plasma/metabolismo , Ratos , Ratos Wistar , Compostos de Sulfidrila/análise , Compostos de Sulfidrila/química , Compostos de Sulfidrila/metabolismo , Ácido Úrico/química , Ácido Úrico/metabolismoRESUMO
A study of several elements of the antioxidative system: Cu-Zn superoxide dismutase (SOD), catalase (CAT), glutathione system (GLU), chemiluminescence (CHE), and antioxidant capacity (AOX), was conducted in 20 demented probable Alzheimer's (DAT), and 15 vascular demented (VD) patients, 19 control (C) subjects, and 11 relatives (F) of one DAT patient. A significant association was found between the variables of the antioxidant system, measured in blood samples, and the neurological pathologies VD and DAT: Kruskal-Wallis test; p = 0.0006 (p = 0.014 when the analysis did not include SOD). This demonstrated that VD and DAT diseases are accompanied by oxidative disorders. The VD and DAT diseases are differentially distinguishable by changes in blood profiles. A graphical method for classification, the Principal Components Analysis (PCA), distinguished between demented and non-demented subjects on the basis of their laboratory variables. A numerical method, Discriminant Functions (DF), constructed to separate the clinical groups on the basis of the same variables, obtained relatively high percentages of success: 92% of demented were detected against healthy subjects; of the latter 82% have been correctly identified as non-demented. Discrimination between VD and DAT patients was achieved for 100% of VD and 86% of DAT patients. DF were similarly successful in detecting the healthy condition of DAT relatives. Possible different mechanisms involved in H2O2 elimination in DAT and VD patients are proposed, where CAT is the responsible enzyme of this reaction in DAT patients, while in VD this function would be achieved mainly through the action of GLU. It seems that SOD levels are stable, at least, within one year. Variations appear to be linked with clinical changes.
Assuntos
Doença de Alzheimer/sangue , Doença de Alzheimer/enzimologia , Antioxidantes/metabolismo , Doenças Vasculares/sangue , Doenças Vasculares/enzimologia , Idoso , Doença de Alzheimer/diagnóstico , Biomarcadores , Catalase/sangue , Interpretação Estatística de Dados , Diagnóstico Diferencial , Eritrócitos/enzimologia , Saúde da Família , Radicais Livres , Glutationa/metabolismo , Humanos , Medições Luminescentes , Estresse Oxidativo/fisiologia , Superóxido Dismutase/sangue , Doenças Vasculares/diagnósticoRESUMO
The oxidative stress in human erythrocytes was studied in asymptomatic and symptomatic patients infected by the human immunodeficiency virus (HIV), and patients with the acquired immunodeficiency syndrome (AIDS). tert-Butyl hydroperoxide initiated chemiluminescence, superoxide dismutase and catalase activities, and total glutathione were evaluated in the erythrocytes and the total antioxidant capacity in the plasma of control, patients infected with HIV that have not yet developed acquired immunodeficiency syndrome, and patients in the later stage of AIDS. tert-Butyl hydroperoxide initiated chemiluminescence was increased by 33% in asymptomatic (stage A1) and symptomatic patients (stage B2) infected with HIV and 82% for patients with AIDS (stage B3) (P < 0.05). While catalase activity did not show any difference between patients and controls, other indices showed differences that, in some cases, reached statistical significance. Superoxide dismutase activity was increased by 24% in stages A1 and B2 of HIV infection and 65% in patients in stage B3 (P < 0.05). Glutathione was decreased by 20% in stages A1 and B2, and by 32% in stage B3 patients (P < 0.05). Total plasma antioxidant capacity was increased in 30 and 57% for the asymptomatic and AIDS patients groups, respectively (P < 0.05). The data indicate that erythrocyte's oxidative stress is associated with the progressive development of HIV disease. Parameters indicating oxidative stress could be an interesting form to screen the evolution of these patients and their response to anti-oxidant therapies.
Assuntos
Eritrócitos/metabolismo , Infecções por HIV/metabolismo , Estresse Oxidativo/fisiologia , Adulto , Antioxidantes/metabolismo , Catalase/sangue , Catalase/metabolismo , Feminino , Radicais Livres/metabolismo , Glutationa/sangue , Glutationa/metabolismo , Humanos , Medições Luminescentes , Masculino , Peróxidos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/sangue , Superóxido Dismutase/metabolismo , terc-Butil HidroperóxidoRESUMO
Rats were made hypertensive by the administration of the nitric oxide synthase inhibitor nitro-L-arginine (LNA, 2.74 mmol/L) in drinking water for 7 d. Hearts from hemodynamically assessed animals were analyzed for lipid peroxidation (LPO), gamma-glutamylcysteine-synthetase (gamma-GCS), glutathione disulfide reductase (GR), glutathione peroxidase (GSHPx), catalase (CAT), superoxide dismutase (SOD), and total radical trapping potential (TRAP) activities. LNA treatment increased the mean arterial blood pressure by 46% and the heart rate by 22% without changing plasma renin activity. LNA treatment resulted in a 30% increase in LPO. gamma-GCS was reduced by 48% and GR by 36% in the cardiac tissue of hypertensive rats as compared to controls. The activity of nonselenium GSHPx was reduced by 27%, and selenium-dependent GSHPx activity in the heart was not affected by LNA treatment. In hypertensive rats, SOD activity was increased by 16%, and CAT was decreased by 46%. TRAP was lower (27%) in the myocardium of hypertensive rats than in that of controls. These data suggest that LNA-induced hypertension is associated with increased myocardial oxidative stress.