RESUMO
Koletsky rats, the genetically obese strain of spontaneously hypertensive rats (SHROB), are the well-accepted animal model of human metabolic syndrome. They are characterized by early onset obesity, spontaneous hypertension, hyperinsulinemia, hyperlipidemia, proteinuria and shortened life-span. One of the factors in the pathogenesis of metabolic syndrome is oxidative stress. The aim of the present study was to compare two parameters related to oxidative stress: the levels of the main intracellular antioxidant, reduced glutathione as well as the indirect indicator of lipid peroxidation damage, thiobarbituric acid-reactive substances (TBARS) in heart, renal cortex and medulla and liver in male lean spontaneously hypertensive rats (SHR) and obese Koletsky rats. We did not find any significant differences in these markers in heart and kidneys. However, we found significantly lower glutathione level in Koletsky rat liver compared with SHR (5.03+/-0.23 vs. 5.83+/-0.14 µmol/g tissue, respectively). On the contrary, we observed significantly higher TBARS levels in Koletsky rat liver compared with SHR (28.56+/-2.15 vs. 21.83+/-1.60 nmol/mg protein, respectively). We conclude that the liver is the most sensitive tissue to oxidative damage with the significantly decreased concentration of glutathione and the significantly increased concentration of TBARS in obese Koletsky rats in comparison with lean control SHR.
Assuntos
Glutationa , Peroxidação de Lipídeos , Fígado , Obesidade , Estresse Oxidativo , Ratos Endogâmicos SHR , Animais , Masculino , Glutationa/metabolismo , Obesidade/metabolismo , Estresse Oxidativo/fisiologia , Ratos , Fígado/metabolismo , Hipertensão/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Rim/metabolismo , Miocárdio/metabolismoRESUMO
Epidemiological studies have demonstrated that n-3 polyunsaturated fatty acid (PUFA) consumption is associated with a reduced risk of atherosclerosis and hyperlipidemia. It is well known that lipid metabolism is also influenced by thyroid hormones. The aim of our study was to test whether n-3 PUFA supplementation (200 mg/kg of body weight/day for 6 weeks given intragastrically) would affect lipid metabolism in Lewis male rats with altered thyroid status. Euthyroid, hypothyroid, and hyperthyroid status of experimental groups was well defined by plasma levels of triiodothyronine, the activity of liver mitochondrial glycerol-3-phosphate dehydrogenase, and by relative heart weight. Fasting blood glucose levels were significantly higher in the hyperthyroid compared to the euthyroid and hypothyroid rats (5.0±0.2 vs. 3.7±0.4 and 4.4±0.2 mmol/l, respectively). In hyperthyroid animals, the concentration of plasma postprandial triglycerides was also increased compared to euthyroid and hypothyroid rats (0.9±0.1 vs. 0.5±0.1 and 0.4±0.1 mmol/l, respectively). On the other hand, hypothyroidism compared to euthyroid and hyperthyroid status was associated with elevated plasma levels of total cholesterol (2.6±0.2 vs. 1.5±0.1 and 1.6±0.1 mmol/l, respectively), LDL cholesterol (0.9±0.1 vs. 0.4±0.1 and 0.2±0.1 mmol/l, respectively) as well as HDL cholesterol (1.6±0.1 vs. 1.0±0.1 and 1.3±0.1 mmol/l, respectively). Supplementation of n-3 PUFA in the present study did not significantly modify either relative heart weight or glucose and lipid levels in any thyroid status.
Assuntos
Ácidos Graxos Ômega-3/metabolismo , Hipertireoidismo/metabolismo , Hipotireoidismo/metabolismo , Metabolismo dos Lipídeos , Animais , Colesterol/metabolismo , Suplementos Nutricionais/análise , Ácidos Graxos Ômega-3/administração & dosagem , Glicerolfosfato Desidrogenase/metabolismo , Humanos , Hipertireoidismo/enzimologia , Hipotireoidismo/enzimologia , Fígado/metabolismo , Masculino , Mitocôndrias/enzimologia , Mitocôndrias/metabolismo , Ratos , Ratos Endogâmicos Lew , Hormônios Tireóideos/metabolismoRESUMO
List of changes: On the basis of author's request the publisher of Physiological Research decided to change the license of the article to CC BY license.
RESUMO
Sex-related cardiovascular differences were observed in humans as well as in experimental animals. Our previous study demonstrated a marked sexual dimorphism in blood pressure (BP) of 9-month-old heterozygous transgenic Ren 2 rats (TGR), in which mouse Ren-2 renin gene was inserted into the genome of normotensive Hannover Sprague-Dawley rats (HanSD). We found significantly elevated BP only in male TGR, whereas BP of TGR females was similar to that of HanSD females. The aim of our present study was to compare BP of 3- and 6-month-old heterozygous TGR with age- and sex-matched HanSD under the same conditions as we measured in 9-month-old rats. We also monitored the amount of oxidative stress marker, thiobarbituric acid-reactive substances (TBARS), and a main intracellular antioxidant, reduced glutathione in the heart, kidneys and liver. We also measured plasma triglycerides and cholesterol levels. We found an increased mean arterial pressure in both female and male 3-month-old TGR (172±17 vs. 187±4 mm Hg, respectively) compared to HanSD (115±5 vs. 133±3 mm Hg, respectively) but there was a marked sexual dimorphism of 6 month-old TGR where only males were hypertensive (145±5 mm Hg) while females became normotensive (123±7 mm Hg). We did not find any relationship between BP values and concentrations of TBARS or glutathione or plasma lipid levels. Our results demonstrated that 6-month-old TGR exhibited a marked sexual BP dimorphism, which was not dependent on the abnormalities in oxidative stress or cholesterol metabolism.
Assuntos
Hipertensão , Renina , Animais , Feminino , Masculino , Ratos , Pressão Sanguínea , Colesterol , Radicais Livres , Glutationa , Rim , Ratos Sprague-Dawley , Ratos Transgênicos , Renina/genética , Substâncias Reativas com Ácido Tiobarbitúrico , Fatores SexuaisRESUMO
In our chronic experiments (over several months), the activity and protein amount of glycerol-3-phosphate dehydrogenase (GPDH) in mitochondria isolated from the liver of adult male and female inbred Lewis strain euthyroid (EU), hyperthyroid (TH), and hypothyroid (HY) rats were analyzed by biochemical and Western blot methods. The TH status was induced by intraperitoneal injections of 3,3',5-triiodo- L-thyronine and the HY status with 0.05% solution of methimazole in drinking water. The TH status led to a significant increase and the HY status to a significant decrease of enzyme activity and protein amount in both male and female animals. These changes were, however, more pronounced in females. The EU and TH female rats also showed a significantly higher activity and the TH female rats showed also a significantly higher enzyme amount in comparison with males, while the HY rats showed low levels in both sexes. The glycerol-3-phosphate-dependent oxygen consumption of freshly isolated rat liver mitochondria from the TH animals was higher in comparison with the EU animals and it was activated by idebenone, a synthetic analogue of coenzyme Q, in both the EU and TH rats. Measurements of serum thyroid hormone levels and analysis of anatomical parameters (relative heart and thyroid gland weights) confirmed that our procedures inducing the TH and HY states are efficient and reliable and that determination of GPDH can serve as an additional criterion for the evaluation of the thyroid hormone status.
Assuntos
Glicerolfosfato Desidrogenase/genética , Hipertireoidismo/metabolismo , Hipotireoidismo/metabolismo , Mitocôndrias Hepáticas/metabolismo , Consumo de Oxigênio , Animais , Modelos Animais de Doenças , Feminino , Expressão Gênica , Glicerolfosfato Desidrogenase/metabolismo , Humanos , Hipertireoidismo/enzimologia , Hipertireoidismo/genética , Hipotireoidismo/enzimologia , Hipotireoidismo/genética , Masculino , Mitocôndrias Hepáticas/enzimologia , Ratos , Ratos Endogâmicos Lew , Hormônios Tireóideos/sangueRESUMO
Coenzyme Q10 (CoQ10), a lipophilic substituted benzoquinone, is present in animal and plant cells. It is endogenously synthetized in every cell and involved in a variety of cellular processes. CoQ10 is an obligatory component of the respiratory chain in inner mitochondrial membrane. In addition, the presence of CoQ10 in all cellular membranes and in blood. It is the only endogenous lipid antioxidant. Moreover, it is an essential factor for uncoupling protein and controls the permeability transition pore in mitochondria. It also participates in extramitochondrial electron transport and controls membrane physicochemical properties. CoQ10 effects on gene expression might affect the overall metabolism. Primary changes in the energetic and antioxidant functions can explain its remedial effects. CoQ10 supplementation is safe and well-tolerated, even at high doses. CoQ10 does not cause any serious adverse effects in humans or experimental animals. New preparations of CoQ10 that are less hydrophobic and structural derivatives, like idebenone and MitoQ, are being developed to increase absorption and tissue distribution. The review aims to summarize clinical and experimental effects of CoQ10 supplementations in some neurological diseases such as migraine, Parkinson´s disease, Huntington´s disease, Alzheimer´s disease, amyotrophic lateral sclerosis, Friedreich´s ataxia or multiple sclerosis. Cardiovascular hypertension was included because of its central mechanisms controlling blood pressure in the brainstem rostral ventrolateral medulla and hypothalamic paraventricular nucleus. In conclusion, it seems reasonable to recommend CoQ10 as adjunct to conventional therapy in some cases. However, sometimes CoQ10 supplementations are more efficient in animal models of diseases than in human patients (e.g. Parkinson´s disease) or rather vague (e.g. Friedreich´s ataxia or amyotrophic lateral sclerosis).
Assuntos
Doenças Mitocondriais , Doenças do Sistema Nervoso , Animais , Antioxidantes/farmacologia , Transporte de Elétrons , Humanos , Mitocôndrias/metabolismo , Doenças Mitocondriais/metabolismo , Doenças do Sistema Nervoso/tratamento farmacológico , Doenças do Sistema Nervoso/metabolismo , Ubiquinona/análogos & derivados , Ubiquinona/uso terapêuticoRESUMO
Sex-related differences were observed not only in human but also in experimental hypertension. The aim of our study was to compare blood pressure (BP) of aged male and female heterozygous transgenic rats (TGR) harboring Ren-2 mouse gene, with their normotensive Hannover Sprague-Dawley (HanSD) controls. At the age of 9 months, systolic (SBP) and diastolic blood pressure (DBP) were measured by a direct puncture of carotid artery in rats awaking from isoflurane anesthesia. Thiobarbituric acid-reactive species (TBARS) formation was monitored as indicator of lipid peroxidation damage in heart, kidney and liver, whereas intracellular content of reduced glutathione was determined in the same organs as the main intracellular antioxidant. Furthermore, plasma triglycerides and total cholesterol as well as high-density lipoprotein (HDL) and low-density lipoprotein (LDL) fractions of cholesterol were measured. As compared to HanSD rats, we found significantly elevated BP only in male TGR (MAP: 123±1 vs. 171±5, SBP: 150±2 vs. 208±7, and DBP: 99±3 vs. 140±4 mm Hg), but not between TGR and HanSD females, which were both normotensive. We also did not find any significant differences in TBARS and reduced glutathione in the three above mentioned organs as well as in plasma cholesterol or its HDL and LDL fractions between transgene-negative HanSD and TGR animals of either sex. However, we found significant sex differences in TBARS, glutathione and plasma lipids in both rat strains. Our results confirmed that aged TGR exhibit a marked sexual BP dimorphism, which does not seem to be dependent on oxidative stress or abnormal cholesterol metabolism.
Assuntos
Envelhecimento/genética , Pressão Sanguínea/fisiologia , Renina/genética , Caracteres Sexuais , Envelhecimento/metabolismo , Animais , Feminino , Masculino , Camundongos , Estresse Oxidativo/fisiologia , Ratos , Ratos Sprague-Dawley , Ratos Transgênicos , Renina/metabolismoRESUMO
The concentration-dependence of tert-butyl hydroperoxide (BHP) inhibitory effect on oxygen consumption in isolated rat liver mitochondria was measured in the presence of various respiratory substrates. Strong inhibitory effect at low concentrations of BHP (15-30 microM) was found for oxoglutarate and palmitoyl carnitine oxidation. Pyruvate and glutamate oxidation was inhibited at higher concentrations of BHP (100-200 microM). Succinate oxidation was not affected even at 3.3 mM BHP. Determination of mitochondrial membrane potential has shown that in the presence of NADH-dependent substrates the membrane potential was dissipated by BHP but was completely restored after addition of succinate. Our data thus indicate that beside peroxidative damage of complex I also various mitochondrial NADH-dependent dehydrogenases are inhibited, but to a different extent and with different kinetics. Our data also show that succinate could be an important nutritional substrate protecting hepatocytes during peroxidative damage.
Assuntos
Mitocôndrias Hepáticas/metabolismo , Estresse Oxidativo , Animais , Respiração Celular , Ácido Glutâmico/metabolismo , Ácidos Cetoglutáricos/metabolismo , Masculino , Potencial da Membrana Mitocondrial , Consumo de Oxigênio , Palmitoilcarnitina/metabolismo , Ácido Pirúvico/metabolismo , Ratos , Ratos Wistar , Ácido Succínico/metabolismo , terc-Butil Hidroperóxido/farmacologiaRESUMO
Mitochondria as an energy generating cell device are very sensitive to oxidative damage. Our previous findings obtained in hepatocytes demonstrated that Complex I of the respiratory chain is more sensitive to oxidative damage than other respiratory chain complexes. We present additional data on isolated mitochondria showing that palmityl carnitine oxidation is strongly depressed at a low (200 microM) tert-butyl hydroperoxide (tBHP) concentration, while oxidation of the flavoprotein-dependent substrate-succinate is not affected and neither is ATP synthesis inhibited by tBHP. In the presence of tBHP, the respiratory control index for palmityl carnitine oxidation is strongly depressed, but when succinate is oxidized the respiratory control index remains unaffected. Our findings thus indicate that flavoprotein-dependent substrates could be an important nutritional factor for the regeneration process in the necrotic liver damaged by oxidative stress.
Assuntos
Mitocôndrias/metabolismo , Oxidantes/farmacologia , Palmitoilcarnitina/metabolismo , Ácido Succínico/metabolismo , terc-Butil Hidroperóxido/farmacologia , Animais , Ácidos Graxos/metabolismo , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Fígado/citologia , Fígado/metabolismo , Masculino , Mitocôndrias/efeitos dos fármacos , NADP/efeitos dos fármacos , NADP/metabolismo , Estresse Oxidativo/fisiologia , Ratos , Ratos WistarRESUMO
Red palm oil (RPO) is a rich natural source of antioxidant vitamins, namely carotenes, tocopherols and tocotrienols. However, it contains approximately 50 % saturated fatty acids the regular consumption of which could negatively modify lipid profile. The aim of our study was to test whether 7 weeks of RPO supplementation (1 g/kg body weight/day) would affect blood glucose and lipid metabolism in adult male Wistar rats with altered thyroid status. We induced hypothyroidism and hyperthyroidism in rats by oral administration of either methimazole or mixture of thyroid hormones. Different thyroid status (EU - euthyroid, HY - hypothyroid and HT - hyperthyroid) was characterized by different serum thyroid hormones levels (total and free thyroxine and triiodothyronine), changes in the activity of a marker enzyme of thyroid status - liver mitochondrial glycerol-3-phosphate dehydrogenase, and altered absolute and relative heart weights. Fasting blood glucose levels were higher in HT rats in comparison with EU and HY rats, but the changes caused by RPO supplementation were not significant. The achievement of the HY status significantly increased serum levels of total cholesterol, as well as with high-density lipoprotein-cholesterol and low-density lipoprotein-cholesterol: 2.43+/-0.15, 1.48+/-0.09, 0.89+/-0.08 mmol/l, compared to EU: 1.14+/-0.06, 0.77+/-0.06, 0.34+/-0.05 mmol/l and HT: 1.01+/-0.06, 0.69+/-0.04, 0.20+/-0.03 mmol/l, respectively. RPO supplementation did not increase significantly levels of blood lipids but tended to increase glutathione levels in the liver. In conclusion, RPO supplementation did not induce the presumed deterioration of glucose and lipid metabolism in rats with three well-characterized alterations in thyroid status.
Assuntos
Glicemia/metabolismo , Suplementos Nutricionais , Lipídeos/sangue , Óleo de Palmeira/farmacologia , Glândula Tireoide/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , Glutationa/metabolismo , Hipertireoidismo/metabolismo , Hipotireoidismo/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Wistar , Hormônios Tireóideos/sangueRESUMO
Our study addresses selected parameters of rat erythrocyte ion transport (Na(+)-K(+) pump, Na(+)-K(+)-2Cl- cotransport, and passive cation fluxes) after acute or chronic hypoxia exposure. We did not find any significant change of ion transport after acute hypoxia. However, chronic hypoxia could modify ion transport because the affinity of Na(+)-K(+) pump for intracellular Na(+) seems to be decreased.
Assuntos
Envelhecimento/metabolismo , Eritrócitos/metabolismo , Hipóxia/metabolismo , Potássio/metabolismo , Sódio/metabolismo , Doença Aguda , Fatores Etários , Altitude , Animais , Doença Crônica , Transporte de Íons , Ratos , Ratos Wistar , Simportadores de Cloreto de Sódio-Potássio/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
Significant relationships between ion transport and membrane lipid composition (cholesterol, total phospholipids and sphingomyelins) were found in erythrocytes of salt hypertensive Dahl rats. In these animals mean cellular hemoglobin content correlated negatively with Na(+)-K(+) pump activity and Na(+) leak but positively with Na(+)-K(+) cotransport activity. Immature erythrocytes exhibit lower mean cellular hemoglobin content (MCHC) than mature ones. The aim of the present study was to find a relationship between erythrocyte maturity, membrane lipid composition and ion transport activity in Wistar rats aged three months which were subjected to repeated hemorrhage (blood loss 2 ml/day for 6 days) to enrich circulating erythrocytes with immature forms. Immature and mature erythrocyte fractions in control and hemorrhaged rats were separated by repeated centrifugation. Hemorrhaged rats had increased number of reticulocytes but reduced hematocrit and MCHC compared to control rats. Immature erythrocytes of hemorrhaged rats differed from mature ones of control animals by elevated Na(+)-K(+) pump activity, reduced Na(+)-K(+) cotransport activity and increased Rb(+) leak. These ion transport changes in immature erythrocytes were accompanied by higher concentration of total phospholipids in their cell membranes. Membrane phospholipid content correlated positively with Na(+)-K(+) pump activity and cation leaks but negatively with Na(+)-K(+) cotransport activity. Moreover, they were also negatively related with MCHC which correlated negatively with Na(+)-K(+) pump activity and Rb(+) leak but positively with Na(+)-K(+) cotransport activity. Thus certain abnormalities of erythrocyte ion transport and membrane lipid composition detected in hypertensive animals might be caused by higher incidence of immature cells.
Assuntos
Membrana Celular/metabolismo , Eritrócitos/metabolismo , Eritropoese/fisiologia , Lipídeos de Membrana/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Animais , Membrana Celular/química , Contagem de Eritrócitos/métodos , Transporte de Íons/fisiologia , Masculino , Lipídeos de Membrana/química , Ratos , Ratos WistarRESUMO
Thyroid hormones are powerful modulators of heart function and susceptibility to arrhythmias via both genomic and non-genomic actions. We aimed to explore expression of electrical coupling protein connexin-43 (Cx43) in the heart of rats with altered thyroid status and impact of omega-3 polyunsaturated fatty acids (omega-3) supplementation. Adult male Lewis rats were divided into following six groups: euthyroid controls, hyperthyroid (treated with T(3)) and hypothyroid (treated with methimazol) with or without six-weeks lasting supplementation with omega-3 (20 mg/100 g/day). Left and right ventricles, septum and atria were used for immunoblotting of Cx43 and protein kinase C (PKC). Total expression of Cx43 and its phosphorylated forms were significantly increased in all heart regions of hypothyroid rats compared to euthyroid controls. In contrast, the total levels of Cx43 and its functional phosphorylated forms were decreased in atria and left ventricle of hyperthyroid rats. In parallel, the expression of PKC epsilon that phosphorylates Cx43, at serine 368, was increased in hypothyroid but decreased in hyperthyroid rat hearts. Omega-3 intake did not significantly affect either Cx43 or PKC epsilon alterations. In conclusion, there is an inverse relationship between expression of cardiac Cx43 and the levels of circulating thyroid hormones. It appears that increased propensity of hyperthyroid while decreased of hypothyroid individuals to malignant arrhythmias may be in part attributed to the changes in myocardial Cx43.
Assuntos
Conexina 43/metabolismo , Ácidos Graxos Ômega-3/farmacologia , Coração/efeitos dos fármacos , Hipertireoidismo/metabolismo , Hipotireoidismo/metabolismo , Proteína Quinase C/metabolismo , Animais , Suplementos Nutricionais , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Miocárdio/enzimologia , Fosforilação , Distribuição Aleatória , Ratos Endogâmicos Lew , Hormônios Tireóideos/sangueRESUMO
The influence of lysophosphatidylcholine (LPC) on H(+)-ATPase, cytochrome oxidase (COX), glycerolphosphate dehydrogenase (GPDH) and malate dehydrogenase (MDH) was followed. The activities of H(+)-ATPase and COX increased with increasing LPC concentration up to 0.5 mg/mg protein when maxima were achieved. This activatory effect is LPC-specific, because Lubrol-treated or frozen-thawed mitochondria showed lower activities of these enzymes. H(+)-ATPase was not influenced by higher concentration of LPC, while COX activity decreased with increasing amount of LPC. The activity of GPDH decreased at very low concentration of LPC and was not further modified at higher LPC concentration. In an attempt to find the concentration of LPC necessary for a complete permeabilization of inner mitochondrial membrane we followed the influence of lysolipid on the release of MDH activity from the mitochondrial matrix. The full activity of this enzyme was obtained with a concentration 0.75 mg LPC/mg protein indicating that mitochondria were completely broken. Our data indicate that LPC significantly affects activity of enzymes connected with mitochondrial membrane and can be useful for evaluation of the importance of phospholipid microenvironment for the enzyme function.
Assuntos
Tecido Adiposo Marrom/enzimologia , Lisofosfatidilcolinas/farmacologia , Mitocôndrias/enzimologia , Tecido Adiposo/enzimologia , Animais , Cricetinae , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Glicerolfosfato Desidrogenase/metabolismo , Membranas Intracelulares/enzimologia , Malato Desidrogenase/metabolismo , Mesocricetus , Mitocôndrias Hepáticas/enzimologia , ATPases Translocadoras de Prótons/metabolismo , Ratos , Ratos EndogâmicosRESUMO
FAD-linked L-glycerol-3-phosphate dehydrogenase purified from liver mitochondria of hyperthyroid rats was incorporated into unilamellar phospholipid liposomes. The incorporation influenced both Vmax,app and Km,app values of the enzyme for its substrate, L-glycerol 3-phosphate. The Km,app for the electron acceptor remained unchanged with a simultaneous slight enhancement of the corresponding Vmax,app value. The steady-state fluorescence anisotropies of the fluorescein isothiocyanate and trimethylammoniumdiphenylhexatriene labels were affected by sodium oleate and calcium ions in the case of both solubilized and liposome-incorporated L-glycerol-3-phosphate dehydrogenase. These results indicate that calcium ions cause a significant alteration of the enzyme conformation. Sodium oleate (used as a model of free fatty acids), besides its direct action on the enzyme itself, affects the enzyme indirectly as well, via alteration of the physical properties of the membrane.
Assuntos
Cálcio/farmacologia , Glicerolfosfato Desidrogenase/metabolismo , Lipossomos/metabolismo , Mitocôndrias Hepáticas/enzimologia , Ácido Oleico , Ácidos Oleicos/farmacologia , Animais , Cardiolipinas/análise , Cardiolipinas/farmacologia , Difenilexatrieno/análogos & derivados , Fluoresceína-5-Isotiocianato , Fluoresceínas , Corantes Fluorescentes , Glicerofosfatos/metabolismo , Hipertireoidismo/enzimologia , Cinética , Lipossomos/análise , Masculino , Fosfolipídeos/análise , Conformação Proteica/efeitos dos fármacos , Ratos , Espectrometria de Fluorescência , TiocianatosRESUMO
The erythrocytes represent an important source of antioxidant capacity of the blood. Catalase (EC 1.11.1.6.) is one of the enzymatic components of their antioxidant defense system. The objective of this study was to follow erythrocyte catalase (CAT) in 7-, 15-, 21-, 35-, 60- and 90-day-old Wistar rats of both sexes in normoxia and after exposure to intensive acute hypobaric hypoxia. During the development CAT activity increases in both sexes, but the rise was usually higher in females. Hypobaric hypoxia increased CAT activity in all studied age groups of both sexes. However, higher CAT activity in females was less affected by hypoxia than the lower activity in males. This was true for nearly all age groups studied. It can be concluded that both ontogenetic aspects and sex differences play a major role in establishing the activity of CAT, which is an important part of the antioxidant defense of the organism.
Assuntos
Envelhecimento/metabolismo , Doença da Altitude/sangue , Doença da Altitude/enzimologia , Catalase/sangue , Eritrócitos/enzimologia , Doença Aguda , Animais , Ativação Enzimática , Feminino , Masculino , Ratos , Ratos Wistar , Fatores SexuaisRESUMO
Hypothalamic paraventricular nucleus (PVN) and rostral ventrolateral medulla (RVLM) play an important role in brain control of blood pressure (BP). One of the important mechanisms involved in the pathogenesis of hypertension is the elevation of reactive oxygen species (ROS) production by nicotine adenine dinucleotide phosphate (NADPH) oxidase. The aim of our present study was to investigate NADPH oxidase-mediated superoxide (O(2)(-)) production and to search for the signs of lipid peroxidation in hypothalamus and medulla oblongata as well as in renal medulla and cortex of hypertensive male rats transgenic for the murine Ren-2 renin gene (Ren-2 TGR) and their age-matched normotensive controls - Hannover Sprague Dawley rats (HanSD). We found no difference in the activity of NADPH oxidase measured as a lucigenin-mediated O(2)(-) production in the hypothalamus and medulla oblongata. However, we observed significantly elevated NADPH oxidase in both renal cortex and medulla of Ren-2 TGR compared with HanSD. Losartan (LOS) treatment (10 mg/kg body weight/day) for 2 months (Ren-2 TGR+LOS) did not change NADPH oxidase-dependent O(2)(-) production in the kidney. We detected significantly elevated indirect markers of lipid peroxidation measured as thiobarbituric acid-reactive substances (TBARS) in Ren-2 TGR, while they were significantly decreased in Ren-2 TGR+LOS. In conclusion, the present study shows increased NADPH oxidase activities in renal cortex and medulla with significantly increased TBARS in renal cortex. No significant changes of NADPH oxidase and markers of lipid peroxidation were detected in the studied brain regions.
Assuntos
Encéfalo/enzimologia , Hipertensão/enzimologia , Rim/enzimologia , Peroxidação de Lipídeos/efeitos dos fármacos , NADPH Oxidases/metabolismo , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Bloqueadores do Receptor Tipo 1 de Angiotensina II/uso terapêutico , Animais , Encéfalo/efeitos dos fármacos , Hipertensão/tratamento farmacológico , Rim/efeitos dos fármacos , Losartan/farmacologia , Losartan/uso terapêutico , Masculino , Distribuição Aleatória , Ratos Transgênicos , Superóxidos/metabolismoRESUMO
Enhanced production of superoxide radicals by nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase in the brain and/or kidney of salt hypertensive Dahl rats has been proposed to participate in the pathogenesis of this form of experimental hypertension. Most information was obtained in young Dahl salt-sensitive (DS) rats subjected to high salt intake prior to sexual maturation. Therefore, the aim of our study was to investigate whether salt hypertension induced in adult DS rats is also accompanied with a more pronounced oxidative stress in the brain or kidney as compared to Dahl salt-resistant (DR) controls. NADPH oxidase activity as well as the content of thiobarbituric acid-reactive substances (TBARS) and conjugated dienes (oxidative index), which indicate a degree of lipid peroxidation, were evaluated in two brain regions (containing either hypothalamic paraventricular nucleus or rostral ventrolateral medulla) as well as in renal medulla and cortex. High salt intake induced hypertension in DS rats but did not modify blood pressure in DR rats. DS and DR rats did not differ in NADPH oxidase-dependent production of ROS, TBARS content or oxidative index in either part of the brain. In addition, high-salt diet did not change significantly any of these brain parameters. In contrast, the enhanced NADPH oxidase-mediated ROS production (without significant signs of increased lipid peroxidation) was detected in the renal medulla of salt hypertensive DS rats. Our findings suggest that there are no signs of enhanced oxidative stress in the brain of adult Dahl rats with salt hypertension induced in adulthood.
Assuntos
Pressão Sanguínea/efeitos dos fármacos , Encéfalo/metabolismo , Hipertensão/metabolismo , Rim/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Cloreto de Sódio na Dieta , Animais , Hipertensão/induzido quimicamente , Rim/efeitos dos fármacos , Masculino , Especificidade de Órgãos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Endogâmicos Dahl , Distribuição TecidualRESUMO
We compared the effect of alpha-tocopheryl succinate (TOS) on succinate-dependent respiration in rat liver mitochondria, homogenate and permeabilized hepatocytes in both a coupled and uncoupled state. In isolated mitochondria, a significant inhibitory effect was observed at a concentration of 5 microM, in liver homogenate at 25 microM and in permeabilized hepatocytes at 50 microM. The inhibitory effect of TOS on succinate respiration in an uncoupled state was less pronounced than in a coupled state in all the experimental models tested. When the concentration dependence of the TOS inhibitory effect was tested, the most sensitive in both states were isolated mitochondria; the most resistant were permeabilized hepatocytes.
Assuntos
Metabolismo Energético/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Mitocôndrias Hepáticas/efeitos dos fármacos , alfa-Tocoferol/farmacologia , Animais , Carbonil Cianeto p-Trifluormetoxifenil Hidrazona/farmacologia , Membrana Celular/metabolismo , Permeabilidade da Membrana Celular , Respiração Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Complexo II de Transporte de Elétrons/metabolismo , Hepatócitos/metabolismo , Masculino , Mitocôndrias Hepáticas/metabolismo , Fosforilação Oxidativa/efeitos dos fármacos , Ratos Wistar , Fatores de Tempo , Desacopladores/farmacologia , alfa-Tocoferol/metabolismoRESUMO
Peroxidation of membrane phospholipids is accompanied by alteration of the structural and functional characteristics of membranes. Lipid peroxidation changes the activities of various enzymes. The present study evaluates the effect of lipid peroxidation on the activity of various ATPases localized on kidney membranes. Our experiments were performed on crude preparation of rat kidney membranes which were exposed to Fe2+.ADP/NADPH-induced lipid peroxidation. The extent of peroxidation was estimated by measuring the thiobarbituric acid-reactive substances. Simultaneously the activities of different ATPases were determined and divided according to their ouabain sensitivity and Mg2+ dependency. We found that 10 min incubation of isolated rat kidney membranes at 37 degrees C with inductors of lipid peroxidation increased the production of thiobarbituric acid-reactive substances from 1.10 +/- 0.26 to 7.72 +/- 2.55 nmol malondialdehyde/mg prot. (+/- SD, n = 4). Under these conditions total ATPase activity was decreased from 681 +/- 77 to 507 +/- 82, ouabain-sensitive Mg(2+)-dependent ATPase (Na+,K(+)-ATPase) activity from 249 +/- 54 to 81 +/- 21 and ouabain-insensitive Mg(2+)-dependent activity from 287 +/- 48 to 173 +/- 58 whereas apparently Mg(2+)-independent ATPase activity was increased from 145 +/- 37 to 253 +/- 42 nmoles P/min/mg prot. (+/- SD, n = 4). The study indicates different mechanisms by which lipoperoxides affect the function of membrane-bound ATPases activities. It is concluded that the ATPases activities are changed during lipid peroxide formation.