RESUMO
Ceramide accumulation in mitochondria has been associated with reperfusion damage, but the underlying mechanisms are not clearly elucidated. In this work we investigate the role of sphingomyelinases in mitochondrial ceramide accumulation, its effect on reactive oxygen species production, as well as on mitochondrial function by using the sphingomyelinase inhibitor, tricyclodecan-9-yl-xanthogenate (D609). Correlation between neutral sphingomyelinase (nSMase) activity and changes in ceramide content were performed in whole tissue and in isolated mitochondria from reperfused hearts. Overall results demonstrated that D609 treatment attenuates cardiac dysfuncion, mitochondrial injury and oxidative stress. Ceramide was accumulated in mitochondria, but not in the microsomal fraction of the ischemic-reperfused (I/R) group. In close association, the activity of nSMase increased, whereas glutathione (GSH) levels diminished in mitochondria after reperfusion. On the other hand, reduction of ceramide levels in mitochondria from I/R+D609 hearts correlated with diminished nSMase activity, coupling of oxidative phosphorylation and with mitochondrial integrity maintenance. These results suggest that mitochondrial nSMase activity contributes to compartmentation and further accumulation of ceramide in mitochondria, deregulating their function during reperfusion.
Assuntos
Ceramidas/metabolismo , Mitocôndrias Cardíacas/metabolismo , Proteínas Mitocondriais/metabolismo , Traumatismo por Reperfusão Miocárdica/metabolismo , Fosforilação Oxidativa , Esfingomielina Fosfodiesterase/metabolismo , Animais , Hidrocarbonetos Aromáticos com Pontes/farmacologia , Glutationa/metabolismo , Proteínas Mitocondriais/antagonistas & inibidores , Traumatismo por Reperfusão Miocárdica/patologia , Norbornanos , Ratos , Esfingomielina Fosfodiesterase/antagonistas & inibidores , Tiocarbamatos , Tionas/farmacologiaRESUMO
Oxidative stress and lung injury induced by short-term exposure to wood smoke were evaluated in guinea pigs through cell profile, bronchoalveolar lavage (BAL), conventional histology and immunohistochemistry (4-hydroxynonenal, 3-nitrotyrosine, Mn-superoxide dismutase, heme oxygenase-1); malondialdehyde and 4-hydroxynonenal concentration, Mn-superoxide dismutase, glutathione reductase, glutathione peroxidase, and catalase activities in plasma, lung and BAL. Total cells increased in BAL, and the percentage of macrophages, neutrophils and lymphocytes augmented (72-96 h). Histopathological examination of lung tissues showed mild thickening of membranous bronchiole walls, infiltration of foamy macrophages and polymorphonuclear leukocytes in bronchial, bronchiolar and intraalveolar spaces. Goblet cell hyperplasia was also observed in bronchial and bronchiolar epithelia. Plasma malondialdehyde concentration was increased at all times, while 4-hydroxynonenal was increased only in plasma and BAL after 24 h. Plasma glutathione reductase activity increased at 24 and 72 h, BAL glutathione peroxidase activity decreased at 72 and 96 h, whereas catalase activity increased in plasma at 72 h, and decreased in BAL at 24 h. Immunostaining intensity to 4-hydroxynonenal, 3-nitrotyrosine, Mn-superoxide dismutase and heme oxygenase-1 was enhanced mainly in macrophages, bronchial/bronchiolar epithelial cells and type II pneumocytes after 72-96 h of wood smoke exposure. Overall, short-term exposure to wood smoke induces alterations in oxidative/antioxidant state in lung and airway injury, similar to those observed in humans with domestic exposure.
Assuntos
Exposição por Inalação/efeitos adversos , Lesão Pulmonar/etiologia , Estresse Oxidativo , Lesão por Inalação de Fumaça/etiologia , Fumaça/efeitos adversos , Madeira , Animais , Antioxidantes/metabolismo , Líquido da Lavagem Broncoalveolar/citologia , Carboxihemoglobina/análise , Contagem de Células , Cobaias , Peróxidos Lipídicos/sangue , Peróxidos Lipídicos/metabolismo , Pulmão/enzimologia , Pulmão/metabolismo , Pulmão/patologia , Lesão Pulmonar/metabolismo , Lesão Pulmonar/patologia , Lesão por Inalação de Fumaça/metabolismo , Lesão por Inalação de Fumaça/patologia , Fatores de TempoRESUMO
Glucosamine (GlcN) is the most used supplement for osteoarthritis treatment. In vitro studies have related GlcN to beneficial and detrimental effects on health. The aim of this study was to evaluate the effects of O-linked-N-acetylglucosaminylation (O-GlcNAc) on GlcN-induced ROS production and Nrf2 expression in human dermal microvascular endothelial cells-1 (HMEC-1) and to evaluate the antioxidant capacity of GlcN compared to well-known antioxidants. For this, we evaluate the antioxidant capacity by in vitro assays. Besides, the GlcN (5-20 mM) effects on cell viability, reactive oxygen species (ROS) production, O-GlcNAc, and nuclear factor erythroid-2-related factor 2 (Nrf2) expression with and without the O-GlcNAc inhibitor OSMI-1 (10 µM) in HMEC-1 were evaluated. GlcN showed high inhibitory concentration (low scavenging activity) against superoxide (O2â¢â, IC20 = 47.67 mM), 2,2-diphenyl-1-picrylhydrazyl (DPPHâ¢, IC50 = 21.32 mM), and hydroxyl (HOâ¢, IC50 = 14.04 mM) radicals without scavenging activity against hydrogen peroxide (H2O2) and low antioxidant capacity determined by oxygen radical absorbance capacity (ORAC, 0.001 mM Trolox equivalent) and ferric reducing antioxidant power (FRAP, 0.046 mM Trolox equivalent). In cell culture, GlcN (20 mM) reduced cell viability up to 26 % and induced an increase in ROS production (up to 70 %), O-GlcNAc (4-fold-higher vs. control), and Nrf2 expression (56 %), which were prevented by OSMI-1. These data suggest an association between O-GlcNAc, ROS production, and Nrf2 expression in HMEC-1 cells stimulated with GlcN.
RESUMO
Cisplatin (CP) is an antineoplastic agent used to treat solid tumors, that has high nephrotoxicity caused by physiologic, hemodynamic, and biochemical alterations. Some studies have shown that naturally derived bioactive compounds in CP-induced nephrotoxicity reduce the side effects of this antineoplastic drug. Pitaya is an endemic fruit from Mexico with a high bioactive compound content, including betalains and phenolic compounds, with reports of antioxidant and anti-inflammatory properties. In this study, the aim was to establish the effect of a pitaya juice concentrate (PJC) on CP-induced nephrotoxicity in Wistar male rats through the identification of metabolites, determination of its chemical composition and antioxidant activity, and evaluation of the protective effect of a PJC on CP-induced nephrotoxicity in rats. The PJC showed a high content of betanins with antioxidant activity by an oxygen radical absorbance capacity assay (1299.6 ± 2.80 Trolox equivalents/g). PJC was administered daily (400 mg day-1, p. o.) for 3 days before CP administration until the end of the experiment. On day four, rats were administered a single injection of CP (6 mg kg, i.p.-1) and sacrificed 72 h later. We observed that CP provoked renal dysfunction (1.0 ± 0.1 vs. 0.4 ± 0.07 serum creatinine levels), oxidative stress, a decrease in nitrate and nitrite (NO2¯/NO3¯) levels (0.1 ± 0.08 vs. 0.4 ± 0.3) and activation of apoptosis and immune responses in kidney tissue. In addition, CP treatment induced tubular damage threefold. PJC administration prevented renal dysfunction (0.5 ± 0.06 vs. 1.0 ± 0.1), normalized degenerative structural damage prevented the increase in lipoperoxidation levels (0.04 ± 0.01 vs. 0.2 ± 0.1) and reduced the apoptosis index by 2.5 in kidney tissue. However, it did not modify the immune response caused by CP. Furthermore, PJC treatment increased nuclear factor erythroid two related factors two protein levels two times and NO2¯/NO3¯ levels 22 times in kidney tissue, which may play a role in the renoprotective effect. In conclusion, the renoprotective effect of PJC on CP-induced nephrotoxicity was associated with the attenuation of dysfunction, structural damage, apoptosis activation, and oxidative stress and was related to changes in the tumor necrosis factor-alpha and renal nitric oxide (NO) pathways. The changes in the NO pathway may be involved in renal hemodynamics. Pitaya could be used as a functional food and therapeutic coadjuvant during CP treatments due to its high bioactive levels and renoprotective compounds.
Assuntos
Antineoplásicos , Nefropatias , Animais , Antineoplásicos/farmacologia , Antioxidantes/metabolismo , Apoptose , Cisplatino/toxicidade , Sucos de Frutas e Vegetais , Nefropatias/induzido quimicamente , Nefropatias/prevenção & controle , Masculino , Óxido Nítrico/metabolismo , Dióxido de Nitrogênio/efeitos adversos , Ratos , Ratos WistarRESUMO
Mast cells (MCs) are important effectors in allergic reactions since they produce a number of pre-formed and de novo synthesized pro-inflammatory compounds in response to the high affinity IgE receptor (FcεRI) crosslinking. IgE/Antigen-dependent degranulation and cytokine synthesis in MCs have been recognized as relevant pharmacological targets for the control of deleterious inflammatory reactions. Despite the relevance of allergic diseases worldwide, efficient pharmacological control of mast cell degranulation has been elusive. In this work, the xanthone jacareubin was isolated from the heartwood of the tropical tree Callophyllum brasilense, and its tridimensional structure was determined for the first time by X-ray diffraction. Also, its effects on the main activation parameters of bone marrow-derived mast cells (BMMCs) were evaluated. Jacareubin inhibited IgE/Ag-induced degranulation in a dose-response manner with an IC50â¯=â¯46â¯nM. It also blocked extracellular calcium influx triggered by IgE/Ag complexes and by the SERCA ATPase inhibitor thapsigargin (Thap). Inhibition of calcium entry correlated with a blockage on the reactive oxygen species (ROS) accumulation. Antioxidant capacity of jacareubin was higher than the showed by α-tocopherol and caffeic acid, but similar to trolox. Jacareubin shown inhibitory actions on xanthine oxidase, but not on NADPH oxidase (NOX) activities. In vivo, jacareubin inhibited passive anaphylactic reactions and TPA-induced edema in mice. Our data demonstrate that jacareubin is a potent natural compound able to inhibit anaphylactic degranualtion in mast cells by blunting FcεRI-induced calcium flux needed for secretion of granule content, and suggest that xanthones could be efficient anti-oxidant, antiallergic, and antiinflammatory molecules.
Assuntos
Anafilaxia/metabolismo , Cálcio/metabolismo , Mastócitos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Receptores de IgE/antagonistas & inibidores , Xantonas/farmacologia , Animais , Degranulação Celular/efeitos dos fármacos , Degranulação Celular/fisiologia , Células Cultivadas , Relação Dose-Resposta a Droga , Líquido Extracelular/efeitos dos fármacos , Líquido Extracelular/metabolismo , Masculino , Mastócitos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Difração de Raios X , Xantonas/isolamento & purificaçãoRESUMO
Oxidative/nitrosative stress is involved in NMDA receptor-mediated excitotoxic brain damage produced by the glutamate analog quinolinic acid. The purpose of this work was to study a possible role of peroxynitrite, a reactive oxygen/nitrogen species, in the course of excitotoxic events evoked by quinolinic acid in the brain. The effects of Fe(TPPS) (5,10,15,20-tetrakis (4-sulfonatophenyl)porphyrinate iron (III)), an iron porphyrinate and putative peroxynitrite decomposition catalyst, were tested on lipid peroxidation and mitochondrial function in brain synaptic vesicles exposed to quinolinic acid, as well as on peroxynitrite formation, nitric oxide synthase and superoxide dismutase activities, lipid peroxidation, caspase-3-like activation, DNA fragmentation, and GABA levels in striatal tissue from rats lesioned by quinolinic acid. Circling behavior was also evaluated. Increasing concentrations of Fe(TPPS) reduced lipid peroxidation and mitochondrial dysfunction induced by quinolinic acid (100 microM) in synaptic vesicles in a concentration-dependent manner (10-800 microM). In addition, Fe(TPPS) (10 mg/kg, i.p.) administered 2 h before the striatal lesions, prevented the formation of peroxynitrite, the increased nitric oxide synthase activity, the decreased superoxide dismutase activity and the increased lipid peroxidation induced by quinolinic acid (240 nmol/microl) 120 min after the toxin infusion. Enhanced caspase-3-like activity and DNA fragmentation were also reduced by the porphyrinate 24 h after the injection of the excitotoxin. Circling behavior from quinolinic acid-treated rats was abolished by Fe(TPPS) six days after quinolinic acid injection, while the striatal levels of GABA, measured one day later, were partially recovered. The protective effects that Fe(TPPS) exerted on quinolinic acid-induced lipid peroxidation and mitochondrial dysfunction in synaptic vesicles suggest a primary action of the porphyrinate as an antioxidant molecule. In vivo findings suggest that the early production of peroxynitrite, altogether with the enhanced risk of superoxide anion (O2*-) and nitric oxide formation (its precursors) induced by quinolinic acid in the striatum, are attenuated by Fe(TPPS) through a recovery in the basal activities of nitric oxide synthase and superoxide dismutase. The porphyrinate-mediated reduction in DNA fragmentation simultaneous to the decrease in caspase-3-like activation from quinolinic acid-lesioned rats suggests a prevention in the risk of peroxynitrite-mediated apoptotic events during the course of excitotoxic damage in the striatum. In summary, the protective effects that Fe(TPPS) exhibited both under in vitro and in vivo conditions support an active role of peroxynitrite and its precursors in the pattern of brain damage elicited by excitotoxic events in the experimental model of Huntington's disease. The neuroprotective mechanisms of Fe(TPPS) are discussed.
Assuntos
Lesões Encefálicas/metabolismo , Doença de Huntington/prevenção & controle , Ácido Peroxinitroso/metabolismo , Porfirinas/uso terapêutico , Radiossensibilizantes/uso terapêutico , Análise de Variância , Animais , Comportamento Animal , Southern Blotting/métodos , Lesões Encefálicas/complicações , Caspase 3 , Caspases/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Interações Medicamentosas , Eletroforese em Gel de Ágar/métodos , Doença de Huntington/etiologia , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/fisiologia , Masculino , Mitocôndrias/fisiologia , Atividade Motora/efeitos dos fármacos , Atividade Motora/fisiologia , Óxido Nítrico Sintase/metabolismo , Ácidos Quinolínicos/farmacologia , Ratos , Ratos Wistar , Teste de Desempenho do Rota-Rod/métodos , Superóxido Dismutase/metabolismo , Vesículas Sinápticas/metabolismo , Sinaptossomos/fisiologia , Tirosina/análogos & derivados , Tirosina/metabolismo , Ácido gama-Aminobutírico/metabolismoRESUMO
Reactive oxygen species have been involved in the pathophysiology of puromycin aminonucleoside (PAN)-nephrosis. The role of H2O2 in these rats may be studied modulating the amount or activity of catalase, which breakdowns H2O2 to water and oxygen. To explore the role of H2O2 in this experimental model, we studied the effect of the in vivo catalase inhibiton with 3-amino-1,2,4-triazole (ATZ) on the course of PAN-nephrosis. Four groups of rats were studied: control rats (CT group), PAN-injected rats (PAN group), ATZ-injected rats (ATZ group), and ATZ- and PAN-injected rats (ATZPAN group). Rats were placed in metabolic cages to collect 24 h urine along the study, ATZ (1 g/kg) was given 24 h before PAN injection (75 mg/kg), and the proteinuria was measured on days 0, 2, 4, 6, 8, and 10. Proteinuria started before (day 4) and was significantly higher on days 6, 8, and 10 in the ATZPAN group than in the PAN group. On day 10, hypercholesterolemia was significantly higher in the ATZPAN group than in the PAN group. These data indicate that the in vivo catalase inhibition magnifies PAN-nephrosis, suggesting that H2O2 is produced in vivo and involved in the renal damage in this experimental disease.
Assuntos
Amitrol (Herbicida)/toxicidade , Catalase/antagonistas & inibidores , Inibidores Enzimáticos/toxicidade , Nefrose/enzimologia , Puromicina Aminonucleosídeo , Animais , Catalase/metabolismo , Progressão da Doença , Avaliação Pré-Clínica de Medicamentos , Glutationa Peroxidase/antagonistas & inibidores , Glutationa Peroxidase/metabolismo , Masculino , Nefrose/induzido quimicamente , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismoRESUMO
Reactive oxygen species are involved in gentamicin (GM) nephrotoxicity, and garlic is effective in preventing or ameliorating oxidative stress. Therefore, the effect of garlic on GM nephrotoxicity was investigated in this work. Four groups of rats were studied: (i) fed normal diet (CT), (ii) treated with GM (GM), (iii) fed 2% garlic diet (GA), and (iv) treated with GM and 2% garlic diet (GM + GA). Rats were placed in metabolic cages and GM nephrotoxicity was induced by injections of GM (75 mg/kg every 12 h) for 6 d. Lipoperoxidation and enzyme determinations were made in renal cortex on day 7. GM nephrotoxicity was made evident on day 7 by (i) tubular histological damage, (ii) enhanced BUN and urinary excretion of N-acetyl-beta-D-glucosaminidase, and (iii) decreased creatinine clearance. These alterations were prevented or ameliorated in GM + GA group. The rise in lipoperoxidation and the decrease in Mn-SOD and glutathione peroxidase (GPx) activities observed in the GM group, were prevented in the GM + GA group. Cu, Zn-SOD activity and Mn-SOD and Cu,Zn-SOD content did not change. CAT activity and content decreased in the GM, GA, and GM + GA groups. CAT mRNA levels decreased in the GM group. The protective effect of garlic is associated with the prevention of the decrease of Mn-SOD and GPx activities and with the rise of lipoperoxidation in renal cortex.
Assuntos
Catalase/metabolismo , Alho , Gentamicinas/toxicidade , Glutationa Peroxidase/metabolismo , Rim/efeitos dos fármacos , Rim/enzimologia , Peroxidação de Lipídeos/efeitos dos fármacos , Plantas Medicinais , Superóxido Dismutase/genética , Acetilglucosaminidase/urina , Animais , Nitrogênio da Ureia Sanguínea , Catalase/genética , Dieta , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Rim/patologia , Córtex Renal/efeitos dos fármacos , Córtex Renal/patologia , Masculino , Estresse Oxidativo , Proteinúria , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Superóxido Dismutase/metabolismoRESUMO
Protein and amino acid metabolism is altered during nephrotic syndrome. However, the expression of the amino acid degrading enzymes has not been well studied. The objective of this work was to assess the expression of hepatic histidase (Hal) and skeletal muscle mitochondrial branched chain amino transferase (BCATm) in rats with experimental nephrotic syndrome induced by a single injection of puromycin aminonucleoside (150 mg/kg). Six days after the injection rats were killed and hepatic Hal and skeletal muscle BCATm activities were measured. Also, total mRNA from both tissues was isolated and Hal and BCATm mRNA expression were analyzed by Northern blot. Rats with NS showed a reduction in food intake with respect to the control group. Hepatic Hal activity increased significantly in nephrotic and pair fed rats by 59% compared to control group. This change in activity was associated with a corresponding increase in Hal mRNA abundance. On the other hand, skeletal muscle BCATm activity and mRNA abundance were similar in the three groups studied. These results suggest that the increase in Hal expression was associated with the reduced food intake and not to the NS. However, BCAT expression did not change indicating the importance of BCAA in body nitrogen conservation.
Assuntos
Regulação Enzimológica da Expressão Gênica , Histidina Amônia-Liase/genética , Fígado/enzimologia , Músculo Esquelético/enzimologia , Síndrome Nefrótica/enzimologia , Síndrome Nefrótica/genética , Transaminases/genética , Animais , Peso Corporal , Modelos Animais de Doenças , Ingestão de Energia , Histidina Amônia-Liase/metabolismo , Humanos , Masculino , Síndrome Nefrótica/induzido quimicamente , Puromicina Aminonucleosídeo , RNA Mensageiro/genética , Ratos , Ratos Wistar , Transaminases/metabolismo , Transcrição GênicaRESUMO
It has been reported that garlic activates nitric oxide synthase in vitro and that chronic inhibition of nitric oxide (NO) synthesis by N omega-nitro-L-arginine-methyl-ester (L-NAME) induces arterial hypertension in rats. In this work, we studied the effect of oral administration of L-NAME for 4 weeks on control and garlic-fed rats. Basal systolic blood pressure was recorded 4 weeks after garlic supplementation, and on weeks 1, 2, 3, and 4 after L-NAME treatment. At the end of the study, the in vivo NO production was evaluated indirectly by measuring the urinary excretion of the stable end products of NO metabolism, nitrite (NO2-) and nitrate (NO3-). It was found that L-NAME induced arterial hypertension on weeks 1-4 in control rats but not in garlic-fed rats, whose blood pressure remained essentially as the basal values. Also, during this time period, blood pressure remained unchanged in garlic-fed rats without L-NAME treatment. Urinary excretion of NO2-/NO3- decreased in L-NAME-treated rats, increased in garlic-fed rats, and remained unchanged in garlic-fed rats treated with L-NAME. It was concluded that garlic blocks the L-NAME-induced hypertension by antagonizing in vivo the inhibitory effect of L-NAME on NO production.
Assuntos
Alho , Hipertensão/prevenção & controle , Óxido Nítrico Sintase/antagonistas & inibidores , Plantas Medicinais , Animais , Pressão Sanguínea/efeitos dos fármacos , Hipertensão/etiologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Nitratos/urina , Nitritos/urina , Ratos , Ratos WistarRESUMO
1. Angiotensin I-converting enzyme (ACE) activity was determined in serum and nine tissues from control and captopril-treated rats, mice, guinea pigs and rabbits. 2. ACE activity was determined with and without sample pretreatment with diamide (total and basal activity, respectively). 3. A very different pattern of response to captopril was observed among the different species. 4. There was no relationship between serum ACE activity and the response to captopril. 5. There were important differences in the determinations of total or basal ACE activities. 6. Endogenous ACE inhibitors were found in some tissues from mouse and rabbit.
Assuntos
Captopril/farmacologia , Peptidil Dipeptidase A/efeitos dos fármacos , Peptidil Dipeptidase A/metabolismo , Animais , Captopril/administração & dosagem , Diamida , Cobaias , Técnicas In Vitro , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Ratos , Ratos Wistar , Especificidade da EspécieRESUMO
Plasma angiotensinogen (Ao) concentration (PAC), urinary Ao excretion (UAE), hepatic levels of Ao mRNA and plasma renin concentration (PRC) were studied in control and nephrotic rats subjected to the following treatments: dexamethasone (DEX), ethinyl-estradiol (EE), tri-iodothyronine (T3), bilateral nephrectomy (NX), captopril (CAP) and adrenalectomy (ADX). In nephrotic rats PAC diminished, UAE and PRC augmented and Ao mRNA levels were not altered. In control rats, DEX, EE, T3 and NX increased PAC and Ao mRNA levels whereas CAP diminished PAC but not affected Ao mRNA. ADX diminished PAC and Ao mRNA levels. In nephrotic rats, these treatments produced the same effect than in control rats except in ADX which did not affect PAC. These data suggest that the decreased PAC is not related to alterations in hepatic Ao gene expression but to elevated PRC and UAE.
Assuntos
Angiotensinogênio/genética , Regulação da Expressão Gênica , Fígado/metabolismo , Síndrome Nefrótica/genética , Adrenalectomia , Angiotensinogênio/sangue , Angiotensinogênio/urina , Animais , Captopril/farmacologia , Dexametasona/farmacologia , Etinilestradiol/farmacologia , Masculino , Nefrectomia , Síndrome Nefrótica/fisiopatologia , RNA Mensageiro/biossíntese , Ratos , Ratos Wistar , Renina/sangue , Tri-Iodotironina/farmacologiaRESUMO
1. Reactive oxygen species are involved in the pathogenesis of puromycin aminonucleoside (PAN) nephrosis and alpha-tocopherol is one of the major anti-oxidants in the body. 2. In the present study, we measured the levels of alpha-tocopherol by high-performance liquid chromatography in the plasma and in nine tissues of control and nephrotic rats obtained 10 days after either 0.9% saline solution or PAN injection, respectively. 3. In nephrotic rats, alpha-tocopherol levels increased four-fold in plasma; however, the molar ratio of alpha-tocopherol/ cholesterol remained unchanged, suggesting that the increase in alpha-tocopherol content was attributable to an increase in plasma lipid concentration. 4. In nephrotic rats, the alpha-tocopherol/cholesterol ratio increased 1.33-fold in adrenal glands and 1.34-fold in the testis, but remained unchanged in heart, spleen, liver, kidney lung, brain and muscle. 5. These data suggest that, in PAN nephrotic rats, there are alterations in the distribution of alpha-tocopherol and there is no deficiency of alpha-tocopherol in plasma or tissues.
Assuntos
Nefrose/metabolismo , Vitamina E/metabolismo , Aldeídos/urina , Animais , Peroxidação de Lipídeos , Masculino , Malondialdeído/urina , Nefrose/induzido quimicamente , Síndrome Nefrótica/metabolismo , Síndrome Nefrótica/patologia , Síndrome Nefrótica/urina , Proteinúria/urina , Puromicina Aminonucleosídeo , Ratos , Ratos Wistar , Fatores de Tempo , Distribuição Tecidual , Vitamina E/sangueRESUMO
1. The concentration of renin and angiotensinogen (Ao) and the activity of angiotensin I-converting enzyme (ACE) was measured in the ascites fluid of nephrotic rats obtained 8 days after puromycin aminonucleoside (PAN) injection. 2. Ascites fluid, serum and urine proteins of these rats were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). 3. Renin, Ao and ACE were found in the ascites fluid and the percentage of the ratio ascites fluid/(plasma or serum) ranged from 5.9 to 9.9%. The electrophoretic analysis revealed that the ascites fluid contained low (Mr < 66 kDa) and high (Mr < 66 kDa) molecular weight proteins. Albumin and six proteins higher than 66 kDa were present both in the ascites fluid and in serum from nephrotic rats. 4. Data from the study suggest that some proteins in the ascites fluid, including renin, Ao and ACE, come from the plasma. It is possible that the loss of renin, Ao and ACE to the ascites fluid may be playing a role in the metabolic alterations of these three proteins in PAN-nephrotic rats.
Assuntos
Líquido Ascítico/química , Síndrome Nefrótica/metabolismo , Proteínas/metabolismo , Sistema Renina-Angiotensina/fisiologia , Angiotensinogênio/metabolismo , Animais , Antibióticos Antineoplásicos , Líquido Ascítico/enzimologia , Eletroforese em Gel de Poliacrilamida , Indicadores e Reagentes , Masculino , Peso Molecular , Síndrome Nefrótica/enzimologia , Síndrome Nefrótica/patologia , Peptidil Dipeptidase A/metabolismo , Proteínas/química , Puromicina Aminonucleosídeo , Radioimunoensaio , Ratos , Ratos WistarRESUMO
Serum and urinary proteins from rats with nephrotic syndrome (NS) induced by puromycin aminonucleoside (PAN) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Analysis was made on days 2, 4, 6, 8, 10, 12, 16, 20, and 30 after PAN injection. Data were compared with control rats (C). Rats developed proteinuria on days 4-30 and hypoproteinemia on days 4-16. Total protein concentration in serum and urine was similar on day 6. SDS-PAGE revealed that urinary albumin augmented on days 4-30 and serum albumin decreased markedly on days 4-20. Albumin concentration in serum and urine was similar on days 4-16. In addition, the study examined serum changes of 7 other proteins (designed as A, B, C, D, E, F, and G) which appeared or increased in urine, and whose molecular weights were higher (A, B, and C) or lower (D, E, F, and G) than that of albumin. In serum, protein A remained unchanged; protein B and G increased; proteins C, D, E, and F decreased. The qualitative pattern of urinary proteins remained essentially unchanged on days 4-30. During the intense proteinuria, the serum concentrations of protein B and albumin were similar and the urine concentrations of proteins C and D became comparable to that found in serum. These 7 serum proteins did not show the same behavior although all of them were excreted in urine. These data indicate that in PAN-nephrotic rats: (a) urinary proteins can be of low and high molecular weight, (b) serum proteins can be regulated independently of their urinary excretion and molecular weight, (c) the urine concentration of total protein and some specific proteins can reach values similar to that found in serum during the intense hypoproteinemia, and (d) the qualitative pattern of urinary proteins was unrelated to the magnitude of proteinuria.
Assuntos
Proteínas Sanguíneas/análise , Síndrome Nefrótica/metabolismo , Proteinúria/metabolismo , Análise de Variância , Animais , Modelos Animais de Doenças , Eletroforese em Gel de Poliacrilamida/métodos , Eletroforese em Gel de Poliacrilamida/estatística & dados numéricos , Hipoproteinemia/metabolismo , Masculino , Peso Molecular , Síndrome Nefrótica/induzido quimicamente , Poliúria/metabolismo , Puromicina Aminonucleosídeo , Ratos , Ratos Wistar , Fatores de TempoRESUMO
Regulation of catalase (CAT) expression, a major antioxidant enzyme that detoxifies H2O2, is very complex. Garlic is effective to prevent or ameliorate oxidative stress probably through its intrinsic antioxidant properties and/or to its ability to modify antioxidant enzyme expression. In this paper we studied the effect of a 2% garlic diet on the renal and hepatic CAT expression (mRNA levels, and enzyme activity, content, synthesis, and degradation). The study was made 2 weeks after feeding rats with a 2% garlic diet. CAT activity and content were measured by a spectrophotometric method and Western blot, respectively. CAT mRNA levels and CAT synthesis (k(s)) and degradation (kD) in vivo were measured by Northern blot and kinetic of reappearance of CAT activity after aminotriazole injection, respectively. Garlic-treatment decreased CAT activity and content, and CAT mRNA levels were unchanged in both tissues. k(s) decreased and kD remained unchanged in kidney and liver. The decrease in k(s) without changes in kD and CAT mRNA levels could explain the low CAT expression in garlic-fed rats. In vivo H2O2 generation in kidney and liver was markedly decreased in garlic-fed rats which could be due to a direct antioxidant effect of garlic. This may be the initial event in the garlic-fed rats that leads to the decreased CAT expression. Our data strongly suggest that the diminished renal and hepatic CAT expression in garlic-fed rats is mediated by post-transcriptional changes (mainly low translational efficiency) which could be an adaptation to the low H2O2.
Assuntos
Catalase/biossíntese , Alho/uso terapêutico , Regulação Enzimológica da Expressão Gênica , Fitoterapia , Plantas Medicinais , Processamento Pós-Transcricional do RNA , Amitrol (Herbicida)/farmacologia , Animais , Antioxidantes/metabolismo , Northern Blotting , Western Blotting , Peso Corporal/efeitos dos fármacos , Creatinina/urina , Comportamento Alimentar/efeitos dos fármacos , Glutationa Peroxidase/biossíntese , Glutationa Peroxidase/sangue , Peróxido de Hidrogênio/metabolismo , Rim/metabolismo , Cinética , Peróxidos Lipídicos/metabolismo , Fígado/metabolismo , Masculino , Biossíntese de Proteínas , RNA/metabolismo , RNA Mensageiro/metabolismo , RNA Ribossômico 18S/metabolismo , Ratos , Ratos Wistar , Espectrofotometria , Superóxido Dismutase/biossíntese , Superóxido Dismutase/sangue , Fatores de TempoRESUMO
Nephrotic syndrome (NS) is characterized by proteinuria, oxidative stress and endogenous hyperlipidemia. Hyperlipidemia and oxidative stress may be involved in coronary heart disease and the progression of renal damage in these patients. Garlic has been suggested to be beneficial in various disease states. Some of the beneficial effects of garlic may be secondary to its hypolipidemic and antioxidant properties. Therefore, the effect of a 2% garlic diet on acute and chronic experimental NS induced by puromycin aminonucleoside (PAN) was studied in this work. Acute NS was induced by a single injection of PAN to rats which were sacrificed 10 days later. Chronic NS was induced by repeated injections of PAN to rats which were sacrificed 84 days after the first injection. Garlic treatment was unable to modify proteinuria in either acute or chronic NS, and hypercholesterolemia and hypertriglyceridemia in acute NS. However, garlic treatment diminished significantly total-cholesterol, LDL-cholesterol and triglycerides, but not HDL-cholesterol in chronic NS. Garlic induced no change in the percentage of sclerotic glomeruli in chronic NS and a significative decrease on the percentage of sclerotic area of these glomeruli (33 +/- 3% in NS+Garlic group vs. 47 +/- 4% in NS group, p = 0.0126). The enhanced in vivo renal H2O2 production and the diminished renal Cu, Zn-SOD and catalase activities in acute NS, and the decreased renal catalase activity in chronic NS were not prevented by garlic treatment. These data indicate that garlic treatment ameliorates hyperlipidemia and renal damage in chronic NS which is unrelated to proteinuria or antioxidant enzymes.