RESUMO
Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant disease characterized by left ventricular hypertrophy (LVH) predominantly affecting the interventricular septum. Cardiac myosin-binding protein C (cMyBP-C) mutations are common causes of FHC. Gene expression profiling was performed in left ventricles of 9-week-old wild-type mice, heterozygous cMyBP-C KO mice displaying asymmetric septal hypertrophy, and homozygous mice developing eccentric LVH. Knocking out one or two cMyBP-C genes leads primarily to gene expression changes indicating an increased energy demand, activation of the JNK and p38 parts of the MAPK pathway and deactivation of the ERK part, and induction of apoptosis. Altered gene expression for processes related to cardiac structure, contractile proteins, and protein turnover was also identified. Many of the changes were more pronounced in the homozygous KO mice. These alterations point to physiological and pathological adaptations in the prehypertrophic heterozygous KO mice and the hypertrophic homozygous mice.
Assuntos
Cardiomiopatia Hipertrófica Familiar/metabolismo , Proteínas de Transporte/metabolismo , Transtornos Cromossômicos/metabolismo , Regulação da Expressão Gênica , Hipertrofia Ventricular Esquerda/metabolismo , Miocárdio/metabolismo , Animais , Apoptose/genética , Cardiomiopatia Hipertrófica Familiar/genética , Cardiomiopatia Hipertrófica Familiar/patologia , Proteínas de Transporte/genética , Transtornos Cromossômicos/genética , Transtornos Cromossômicos/patologia , MAP Quinases Reguladas por Sinal Extracelular , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/genética , Heterozigoto , Homozigoto , Hipertrofia Ventricular Esquerda/genética , Hipertrofia Ventricular Esquerda/patologia , MAP Quinase Quinase 4/genética , MAP Quinase Quinase 4/metabolismo , Sistema de Sinalização das MAP Quinases/genética , Camundongos , Camundongos Knockout , Miocárdio/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Septo Interventricular/metabolismo , Septo Interventricular/patologia , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
The concentration and composition of fatty acids in four lipid classes in biopsies of dog left ventricular myocardium were determined, using gas-liquid chromatography. When precautions were taken to minimize lipolysis during storage of the tissue and the homogenization process, the following results were obtained: 29 +/- 10 nmol non-esterified fatty acids, 2.98 +/- 2.41 mumol triacylglycerol, 149 +/- 51 nmol cholesteryl esters and 23.76 +/- 3.38 mumol phospholipid (expressed as fatty acid moiety per gram of wet tissue). The concentration of non-esterified fatty acids was 15 to 300 times lower than reported in literature. The main constituents of the non-esterified fatty acids were palmitic, stearic and oleic acid. Triacylglycerol consisted of approximately 40% esterified oleic acid. Linoleic acid accounted for 40% of the fatty acids in the cholesteryl-esters class. More than half of the fatty acid moiety of total phospholipids was linoleic acid and arachidonic acid.
Assuntos
Ácidos Graxos/análise , Miocárdio/análise , Animais , Ésteres do Colesterol/análise , Cromatografia Gasosa , Cães , Ácidos Graxos não Esterificados/análise , Feminino , Masculino , Fosfolipídeos/análise , Preservação de Tecido , Triglicerídeos/análiseRESUMO
An enzyme linked immuno-sorbent assay (ELISA) of the sandwich type for the determination of heart-type fatty acid-binding protein (H-FABPc) was developed, making use of the streptavidin-biotin system. The assay turned out to be virtually disturbance insensitive and showed a detection limit for H-FABPc of 0.2 micrograms/l with an intra- and inter-assay variation of 5% and 14%, respectively. The H-FABPc content of adult rat heart muscle was found to be 0.740 +/- 0.120 mg/g wet weight. The H-FABPc content of a number of skeletal muscles varied from 0.013 to 0.303 mg/g wet weight and was related to the content of type I muscle fibers of these tissues, suggesting a role for H-FABPc in intracellular fatty acid metabolism. The assay was further applied to study the release of H-FABPc from isolated rat heart during normoxic Langendorff perfusion, as compared to that of lactate dehydrogenase (LDH), into fluid derived from the right ventricular cavity (Qrv) and that from the interstitial space (Qi). Total release of H-FABPc per 15 min amounted to 0.015 +/- 0.010% but that of LDH to 0.080 +/- 0.040% of their total tissue content. Furthermore, for both H-FABPc and LDH 80% was released into Qi, which only accounted for 1-2% of total flow. These findings suggest that during normoxic perfusion of rat heart H-FABPc, and LDH are released from different cellular compartments and that the bulk amount of released intracellular proteins is transported via the lymph instead of being directly released into the bloodstream.
Assuntos
Proteínas de Transporte/análise , Ácidos Graxos/metabolismo , Miocárdio/química , Proteínas de Neoplasias , Proteínas do Tecido Nervoso , Animais , Proteínas de Bactérias , Biotina , Proteínas de Transporte/metabolismo , Ensaio de Imunoadsorção Enzimática , Proteína 7 de Ligação a Ácidos Graxos , Proteínas de Ligação a Ácido Graxo , Peroxidase do Rábano Silvestre/análise , L-Lactato Desidrogenase/metabolismo , Músculos/química , Perfusão , Coelhos , Ratos , EstreptavidinaRESUMO
Mitochondrial oxidation of long-chain fatty acids provides the majority of the energy required for cardiac functioning. Several proteins, including the integral membrane protein FATP (Fatty Acid-Transport Protein), are being implicated in the process of myocardial fatty acid uptake. To further characterize the role of FATP in rat myocardial fatty acid utilization, cDNA encoding rat FATP was cloned. The inferred amino acid sequence indicates that rat FATP is highly homologous (97%) with its murine equivalent. Moreover, rodent FATPs share several well-conserved regions with putative counterparts found in yeast and nematode. Given the large evolutionary distance between these species, these regions might be important for protein function. The predicted membrane topology of rat FATP is discussed.
Assuntos
Proteínas de Transporte/genética , Ácidos Graxos/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana Transportadoras , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Clonagem Molecular , DNA Complementar , Evolução Molecular , Proteínas de Transporte de Ácido Graxo , Dados de Sequência Molecular , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Homologia de Sequência de AminoácidosRESUMO
The purpose of the present work was to study the acute regulation of glucose uptake in cultured cardiac endothelial cells (CEC). Two types of potential stimuli were considered: (1) agents that are known to acutely stimulate glucose transport (i.e., within minutes) in fat and muscle tissues and (2) agents that influence endothelial cell function. Among the former agents, neither insulin, nor catecholamines (adrenaline, dopamine, phenylephrine), nor serotonin affected the rate of glucose transport in CEC, while SH-group reagents (phenylarsine oxide, diamide or menadione) were inhibitory. Among the factors of the second group that were tested (heparin, ADP, histamine, bradykinin), histamine was found to stimulate glucose transport in CEC by 10-50%. This effect was concentration-dependent (with an EC50 value approximately equal to 12 microM) and reached a maximum within 5 min upon histamine addition. This stimulation of glucose transport was suppressed by pyrilamine (100 nM), a specific H1-receptor antagonist, but not by cimetidine (100 microM), a H2-selective antagonist. Northern blot and Western blot analysis of CEC extracts revealed the presence of the ubiquitous glucose transporter isoform GLUT1 mRNA and protein, but not of the 'insulin-regulatable' isoform GLUT4. In conclusion, this is the first report on an acute stimulation of glucose transport in cardiac endothelial cells, in particular, and in an insulin-unresponsive cell type, in general. The effect of histamine is most likely mediated by H1-receptors and cannot be accounted for by a recruitment of GLUT4.
Assuntos
Endotélio Vascular/metabolismo , Glucose/metabolismo , Histamina/farmacologia , Proteínas Musculares , Difosfato de Adenosina/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Células Cultivadas , Cimetidina/farmacologia , Vasos Coronários/citologia , Desoxiglucose/metabolismo , Transportador de Glucose Tipo 1 , Transportador de Glucose Tipo 4 , Insulina/farmacologia , Masculino , Proteínas de Transporte de Monossacarídeos/biossíntese , Pirilamina/farmacologia , RNA Mensageiro/biossíntese , Ratos , Compostos de SulfidrilaRESUMO
The effect of Ca2+ in vitro on pregnenolone production rates under various incubation conditions by mitochondrial fractions isolated from testes of normal rats and of rats after in vivo treatment with luteinizing hormone has been investigated. Concentrations of Ca2+ in the range of 0.1-0.5 mM stimulated succinate supported pregnenolone production in mitochondrial fractions from both control and luteinizing hormone treated testes. When mitochondrial fractions were isolated in 0.25 M sucrose without additions, Ca2+ in vitro increased succinate supported pregnenolone production rates in mitochondrial fractions isolated from control testes to a greater extent than in mitochondrial fractions, from luteinizing hormone treated testes. Production rates in control mitochondrial fraction, incubated in the presence of initial Ca2+ concentrations of 0.7 mM and higher were almost similar to production rates in relevant luteinizing hormone treated mitochondria.
Assuntos
Cálcio/farmacologia , Hormônio Luteinizante/farmacologia , Mitocôndrias/metabolismo , Pregnenolona/biossíntese , Rutênio Vermelho/farmacologia , Rutênio/farmacologia , Testículo/metabolismo , Envelhecimento , Animais , Ácido Edético/farmacologia , Cinética , Masculino , Mitocôndrias/efeitos dos fármacos , NADP/farmacologia , Ratos , Testículo/efeitos dos fármacos , Testículo/crescimento & desenvolvimentoRESUMO
Steroid production and enzyme activities were examined in preparations of whole testis tissue, isolated interstitial tissue and seminiferous tubules obtained from adult rats with intact pituitaries receiving daily subcutaneous injections of 100 I.U. human chorionic gonadotropin for 5 days and from control animals. After human chorionic gonadotropin administration testosterone concentrations were increased in total homogenates of whole testis tissue, interstitial tissue and seminiferous tubules. The testosterone production from endogenous precursors was enhanced only in total homogenates of whole testis tissue and interstitial tissue obtained from testes of human chorionic gonadotropin-treated rats. The production of testosterone in the corresponding homogenates of isolated seminiferous tubules was very low. The specific activity of 3 beta-hydroxysteroid dehydrogenase was increased in total homogenates of whole testis tissue, isolated interstitial tissue and seminiferous tubules. No effect was observed on the specific activities of marker enzymes such as cytochrome c oxidase, monoamine oxidase, steroid sulfatase and lactate dehydrogenase, whereas the specific activities of carboxyl esterase were decreased in homogenates of whole testis tissue and interstitial tissue. Total activity of monoamine oxidase was increased in homogenates of interstitial tissue of tests from human chorionic gonadotropin treated rats. After the same prolonged human chorionic gonadotropin treatment the concentration of pregnenolone was increased in mitochondrial fractions of whole testis tissue, interstitial tissue and seminiferous tubules, and the amount of protein isolated in the mitochondrial fraction of interstitial tissue increased by 40%. Steroid production (estimated as pregnenolone) from endogenous precusors by mitochondrial fractions of whole testis tissue and interstitial tissue were increased after human chorionic gonadotropin treatment, for whole testis from 580 pmol/mg mitochondrial protein per h to 1420 pmol/mg per h; and for interstitial tissue from 2665 pmol/mg per h to 7050 pmol/mg per h. The production of pregnenolone in mitochondrial fractions obtaine from isolated seminiferous tubules was very low and contributed hardly at all to the total pregnenolone production in mitochondrial fractions of whole testis tissue from normal rats as well as from human chorionic gonadotropin-treated rats.
Assuntos
Gonadotropina Coriônica/farmacologia , Mitocôndrias/metabolismo , Pregnenolona/biossíntese , Testículo/metabolismo , Testosterona/biossíntese , Animais , Ácidos Carboxílicos , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Esterases/metabolismo , Humanos , Hidroxiesteroide Desidrogenases/metabolismo , L-Lactato Desidrogenase/metabolismo , Masculino , Monoaminoxidase/metabolismo , Tamanho do Órgão , Proteínas/metabolismo , Ratos , Sulfatases/metabolismo , Fatores de TempoRESUMO
An attempt has been made to correlate the rapid effect of luteinizing hormone on testicular steroid production in vivo with testicular steroid concentrations and in vitro steroid production rates in testis tissue preparations. Within 20 min after intravenous administration of 25 mug luteinizing hormone, increases were observed in testosterone concentrations in testicular venous plasma and in whole testis tissue and in pregnenlone concentrations isolated testis mitochondrial fractions. Testosterone production by whole testis homogenates and pregnenolone production by isolated mitochondrial fractions were significantly increased within 5 min after in vivo administration of luteinizing hormone. Injection of cycloheximide 10 min prior to luteinizing hormone prevented the stimulating effect of luteinizing hormone to steroid levels in testicular venous plasma and testis tissue and on steroid production rates by preparations of rat testis tissue. Cycloheximide treatment of control animals did not significantly alter testosterone concentrations and testosterone production rates vitro, although mitochondrial pregnenolone concentrations and production rates were decreased. Testosterone production by whole testis homogenates as well as the pregnenolone production by isolated mitochondrial fractions obtained from luteinizing hormone treated testes and control glands showed a biphasic time curve A period (5-10 min) of high steroid production was followed by a period lower steroid production. Addition of 25 mug luteinizing hormone or 10(-8)--10(-5) M adenosine 3':5'-monophosphate (cyclic AMP) to the incubation medium had no effect pregnenolone production by isolated mitochondrial fractions. Administration of leuteinizing hormone in vivo markedly enhance the stimulating effect of Ca2+ on testosterone production by whole testis homogenates and on pregnenolone production by isolated mitochondrial fractions.
Assuntos
Cálcio/farmacologia , Cicloeximida/farmacologia , Hormônio Luteinizante/farmacologia , Esteroides/biossíntese , Testículo/metabolismo , Animais , Sistema Livre de Células , Hidroxiesteroide Desidrogenases/metabolismo , Masculino , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Pregnenolona/biossíntese , Ratos , Testículo/efeitos dos fármacos , Testosterona/biossíntese , Fatores de TempoRESUMO
In the three species under investigation (dog, rat and man) a gradient from serum to heart tissue for total non-esterified fatty acids was assessed. The ratios serum/left ventricular tissue in dogs, serum/right auricular appendage in dogs, serum/whole heart tissue in rats and serum/right auricular appendage in man were found to be 6.4, 2.5, 5.6 and 2.8, respectively. The highest gradient was found for oleic acid, whereas no significant gradient for arachidonic acid could be detected. In the dog the arterio:local venous differences of non-esterified fatty acids across the left ventricular tissue correlated better with the serum/tissue ratio of non-esterified fatty acids than with the arterial non-esterified fatty acid level. Since the correlation coefficient (0.74) was still far from excellent, more factors than the non-esterified fatty acid serum/tissue gradient are likely to be involved in determining the extent to which non-esterified fatty acids are extracted by myocardial tissue.
Assuntos
Ácidos Graxos não Esterificados/metabolismo , Miocárdio/metabolismo , Animais , Cães , Ácidos Graxos não Esterificados/sangue , Ácidos Graxos não Esterificados/isolamento & purificação , Ventrículos do Coração/metabolismo , Humanos , Especificidade de Órgãos , Ratos , Especificidade da Espécie , Relação Estrutura-AtividadeRESUMO
To study the influence of membrane fatty acid composition on the formation of prostanoids and hydroxy fatty acids by rat peritoneal mast cells (MC), animals were fed three different types of fatty acids: mackerel oil (MO), abundant in n-3 fatty acids; sunflower seed oil (SO), rich in linoleic acid; and hydrogenated coconut oil (HCO), mainly containing saturated fatty acids. The presence of n-3 fatty acids in the diet resulted in the incorporation of 20:5(n-3), 22:5(n-3) and 22:6(n-3) in MC phospholipids. A decrease of arachidonic acid, 20:4(n-6), was observed in MC-phospholipids of the MO-fed animals. Furthermore, increasing the relative amounts of 18:2(n-6) in the diet (SO group) led to an increased incorporation of linoleic acid, 18:2(n-6) in MC phospholipids when compared to both other dietary groups. The changes in MC phospholipid fatty acid composition were (partly) reflected in the formation of prostanoids and hydroxy fatty acids upon stimulation with the calcium ionophore A23187. The decrease in arachidonic acid content in MC phospholipids of MO-fed rats resulted in a decreased formation of PGD2 when compared to both other groups. Also, the increased amounts of 18:2(n-6) in MC phospholipids of SO-fed rats resulted in an increased formation of 9- and 13-HODE upon stimulation. The results show that modifications in the fatty acid composition of the diet influences MC membrane fatty acid composition which ultimately results in changes in prostanoid and hydroxy fatty acid synthesis by MC upon stimulation with the calcium ionophore A23187.
Assuntos
Gorduras na Dieta/farmacologia , Ácidos Graxos/biossíntese , Mastócitos/efeitos dos fármacos , Prostaglandinas/biossíntese , Animais , Ácido Araquidônico/metabolismo , Peso Corporal , Calcimicina , Membrana Celular/metabolismo , Óleo de Coco , Óleos de Peixe/farmacologia , Ácido Linoleico , Ácidos Linoleicos/metabolismo , Masculino , Mastócitos/metabolismo , Cavidade Peritoneal , Fosfolipídeos/metabolismo , Óleos de Plantas/farmacologia , Ratos , Ratos Wistar , Óleo de GirassolRESUMO
The release of cardiac fatty acid-binding protein (cFABP) and of fatty acids from isolated rat hearts was measured during both reperfusion following 60 min of ischemia and the calcium paradox (readmission of Ca2+ after a period of Ca2+-free perfusion). Total cFABP release was much more pronounced after Ca2+ readmission (over 50% of tissue content) than during post-ischemic reperfusion (on average, 3% of tissue content), but in both cases, it closely paralleled the release of lactate dehydrogenase. Only minor amounts of long-chain fatty acids, if any, were released from the heart. These observations are challenging the idea that cFABP plays a fatty acid-buffering role under the pathophysiological conditions studied.
Assuntos
Cálcio/fisiologia , Proteínas de Transporte/metabolismo , Miocárdio/metabolismo , Proteínas de Neoplasias , Proteínas do Tecido Nervoso , Animais , Cálcio/farmacologia , Doença das Coronárias/metabolismo , Proteína 7 de Ligação a Ácidos Graxos , Proteínas de Ligação a Ácido Graxo , Coração/efeitos dos fármacos , Técnicas In Vitro , Cinética , L-Lactato Desidrogenase/metabolismo , Masculino , Modelos Biológicos , Perfusão , Ratos , Ratos Endogâmicos LewRESUMO
The addition of lactate (5.6 mM) to a perfusion medium containing glucose (11 mM) stimulated the turnover of the cardiac triacylglycerol pool throughout the perfusion period as indicated by increased glycerol release in association with maintained levels of triacylglycerols. Attenuation of feedback inhibition of triacylglycerol lipase by fatty acids as a possible cause of the elevated triacylglycerol turnover rate should be ruled out, since tissue fatty acid levels were 3-times higher in glucose plus lactate perfused hearts than in hearts perfused with glucose as the sole substrate. The present findings are in favor of the notion that lactate enhances triacylglycerol turnover through increased glycerol 3-phosphate levels.
Assuntos
Coração/efeitos dos fármacos , Lactatos/farmacologia , Miocárdio/metabolismo , Triglicerídeos/metabolismo , Animais , Ácidos Graxos/análise , Glicogênio/análise , Hemodinâmica , Técnicas In Vitro , Masculino , Ratos , Ratos Endogâmicos LewRESUMO
We studied how the release of histamine and prostaglandin D2 (PGD2) were connected in stimulated rat peritoneal mast cells, and to what extent these processes were controlled by the cytosolic Ca2+ concentration, [Ca2+]i, and protein tyrosine kinases. In the presence of 1 mM CaCl2, the G-protein activating compound 48/80 (10 micrograms/ml) evoked a transient rise in [Ca2+]i and a relatively high secretion of histamine, but only a low release of PGD2. In contrast, 5 microM thapsigargin (an inhibitor of endomembrane Ca(2+)-ATPases) and 5 microM ionomycin evoked high and prolonged rises in [Ca2+]i, and stimulated the cells to release relatively small amounts of histamine and high amounts of PGD2. Stimulation of the cells with CaCl2 and 10 microM ATP4- gave only minor quantities of histamine and PGD2, despite of the micromolar level of [Ca2+]i reached. When CaCl2 was replaced by EGTA, rises in [Ca2+]i as well as release of histamine and PGD2 were reduced with each agonist, but the preference of agonists to release more histamine or PGD2 remained unchanged. In mast cells with depleted Ca2+ stores, the addition of CaCl2 stimulated the store-regulated Ca2+ entry resulting in a prolonged rise in [Ca2+]i. However, simultaneous addition of compound 48/80 and CaCl2 was required for release of histamine and PGD2. In cells with full stores, PGD2 release evoked by compound 48/80 was greatly reduced by genistein and methyl-2,5-dihydroxycinnamate, two structurally unrelated inhibitors of protein tyrosine kinases, whereas histamine secretion was not influenced by these inhibitors. Similarly, with thapsigargin or ionomycin as agonist, PGD2 release was more sensitive to the tyrosine kinase inhibitors than histamine secretion. We conclude that in activated rat peritoneal mast cells: (i) the influx of extracellular Ca2+ potentiates agonist-evoked rises in [Ca2+]i as well as histamine secretion and PGD2 release; (ii) the amplitude of the [Ca2+]i rise does not determine the preferential effect of agonists to release more histamine or more PGD2; (iii) the relatively high PGD2 release evoked by thapsigargin and ionomycin is probably due to their potency to evoke a prolonged rise in [Ca2+]i and to activate protein tyrosine kinases.
Assuntos
Cálcio/metabolismo , Histamina/metabolismo , Peritônio/metabolismo , Prostaglandina D2/metabolismo , Proteínas Tirosina Quinases/metabolismo , Animais , Citosol/metabolismo , Masculino , Mastócitos/metabolismo , Proteínas Tirosina Quinases/antagonistas & inibidores , Ratos , Ratos Wistar , Terpenos/farmacologia , Tapsigargina , p-Metoxi-N-metilfenetilamina/farmacologiaRESUMO
OBJECTIVE: The effect of exogenous lactate and pyruvate on the recovery of coronary flow (total, regional) after ischaemia as a function of the duration of ischaemia was evaluated. METHODS: Isolated, ejecting rat hearts were subjected to ischaemia for 15, 30, or 45 minutes. Glucose (11 mM) was present as the basal substrate in the perfusion medium and lactate (5 mM) or pyruvate (5 mM) was added as the cosubstrate. Flow variables were measured by the timed collection of coronary effluents and by the radioactive microsphere technique. RESULTS: In the lactate perfused hearts, reactive hyperaemia was present after 15 minutes but absent after 30 and 45 minutes of ischaemia. Total coronary flow was significantly reduced after 45 minutes of ischaemia. Transmural flow was impaired after 15, 30, and 45 minutes of ischaemia in the lactate perfused hearts, -that is, flow in the inner layers of the left ventricle was transiently reduced after 15 minutes and remained continuously depressed after 30 and 45 minutes of ischaemia. The pyruvate perfused hearts showed reactive hyperaemia after 15 and 30 minutes of ischaemia. After 45 minutes total coronary flow was reduced below the value before ischaemia, and in particular, the inner layers of the left ventricle were severely deprived of flow as in lactate perfused hearts. When neither lactate nor pyruvate was added to the perfusion medium (containing glucose), impairment of coronary flow in the inner layers of the left ventricle was only transiently obvious after 30 and 45 minutes of ischaemia. Impaired perfusion of the inner layers during reperfusion resulted in delayed washout of lactate dehydrogenase. CONCLUSIONS: Exogenous substrates modify the recovery of flow after ischaemia. In the presence of exogenous lactate, severe disturbances of flow are already obvious after 30 minutes of ischaemia in the inner layers of the left ventricle. Exogenous pyruvate delays impairment of flow in the inner layers compared with exogenous lactate.
Assuntos
Circulação Coronária/efeitos dos fármacos , Lactatos/farmacologia , Isquemia Miocárdica/fisiopatologia , Piruvatos/farmacologia , Animais , Glucose/farmacologia , Técnicas In Vitro , L-Lactato Desidrogenase/sangue , Ácido Láctico , Masculino , Reperfusão Miocárdica , Ácido Pirúvico , Ratos , Ratos Endogâmicos Lew , Fatores de TempoRESUMO
Fatty acids are important energy donors for the healthy heart. These substrates are supplied to the myocardium bound to albumin to overcome their low solubility in aqueous solutions such as blood plasma. Transport from the microvascular compartment to the mitochondria inside the cardiomyocytes is most likely a combination of passive and protein-mediated diffusion. Alterations in tissue content of fatty acid-transport proteins may contribute to myocardial diseases such as the diabetic heart, and cardiac hypertrophy and failure.
Assuntos
Cardiomiopatias/metabolismo , Ácidos Graxos/metabolismo , Miocárdio/metabolismo , Trifosfato de Adenosina/genética , Trifosfato de Adenosina/metabolismo , Animais , Transporte Biológico , Cardiomegalia/genética , Cardiomegalia/metabolismo , Cardiomiopatias/genética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Diabetes Mellitus/genética , Diabetes Mellitus/metabolismo , Regulação da Expressão Gênica , Humanos , Mitocôndrias Cardíacas/metabolismoRESUMO
Isolated working hearts of 16 month old spontaneously hypertensive rats (SHR, n = 8) and age matched Wistar-Kyoto (WKY, n = 8) rats were exposed to 30 min global normothermic ischaemia followed by 60 min reperfusion. The hearts were routinely perfused at an afterload level of 13.3 kPa and a preload level of 1.0 kPa. The control values of left ventricular pressure, its maximal positive first derivative (dP1v/dtmax), coronary flow per gram heart tissue, and release of lactate and enzymes such as lactate dehydrogenase and aspartate aminotransferase were comparable in both groups. WKY rat hearts ejected almost twice as much perfusate per gram heart weight as the SHR hearts. In pressure-flow curves, obtained during the control period in SHR hearts, cardiac output was independent of changes in afterload, varying between 10.7 and 18.7 kPa. In contrast, in WKY rat hearts increases in afterload resulted in a progressive decrease in cardiac output. Reperfusion of the SHR hearts after 30 min of global normothermic ischaemia resulted in a poor recovery of cardiac output (13% of the control values) and dP1v/dtmax (32%) compared with the values in the WKY rat hearts (66% and 91% of the control values respectively). Reactive hyperaemia was prominent in the WKY rat hearts but completely absent in the SHR hearts. During one hour reperfusion, SHR hearts lost 3.5 times more lactate dehydrogenase and 2.5 times more aspartate aminotransferase than the WKY rat hearts. Pressure-flow curves, obtained during the reperfusion period, showed modest recovery of myocardial function of the WKY rat hearts at the lowest afterload level tested but completely depressed myocardial function of the SHR hearts at all afterload levels. Heart tissue contents of adenosine triphosphate and creatine phosphate after one hour of reperfusion were lower in the SHR than in the WKY rats, but compared with native values a comparable percentage decrease was seen in both groups of rats.
Assuntos
Vasos Coronários , Coração/fisiopatologia , Hipertensão/fisiopatologia , Isquemia/fisiopatologia , Trifosfato de Adenosina/metabolismo , Animais , Aspartato Aminotransferases/metabolismo , Débito Cardíaco , Cardiomegalia/fisiopatologia , L-Lactato Desidrogenase/metabolismo , Miocárdio/metabolismo , Perfusão , Fosfocreatina/metabolismo , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKYRESUMO
OBJECTIVE: The aim was to examine differences between the postischaemic functional and biochemical recovery of adult and aged hypertrophied hearts. METHODS: Isolated hypertrophied hearts of adult and aged spontaneously hypertensive rats (SHRadult; SHRaged) and normal hearts of age matched Wistar-Kyoto rats (WKYadult; WKYaged) were perfused in an ejecting heart preparation. Haemodynamic function was monitored before and after 45 min of ischaemia. Coronary effluent samples and tissue biopsies were taken for biochemical analysis. RESULTS: After ischaemia, in SHRadult and WKYadult the maximum positive first derivative of the left ventricular pressure (dP/dtmax) was restored to 105% and 97% respectively of the preischaemic values. Left ventricular developed pressure recovered to 80% (SHRadult) and 97% (WKYadult), while cardiac output reached 71% (SHRadult) and 99% (WKYadult) of preischaemic levels. In SHRaged and WKYaged the dP/dtmax recovered to 26% and 60% respectively (both p < 0.05 compared to the preischaemic values). The left ventricular developed pressure recovered to 36% in SHRaged and to 73% in WKYaged (both p < 0.05), while cardiac output was restored to 6% in SHRaged and 38% in WKYaged (both p < 0.05). Throughout reperfusion, left ventricular end diastolic pressure remained significantly elevated in SHRaged, and was associated with a prominent subendocardial underperfusion, suggesting an impaired diastolic functional recovery. Overall haemodynamic recovery was significantly better in the WKYaged than in the SHRaged. The preischaemic total adenine nucleotides content was comparable in all groups, but creatine phosphate levels were significantly lower in both aged groups than in adult groups. In all but the WKYadult, the total adenine nucleotides were depressed upon reperfusion, while creatine phosphate normalised, except in SHRaged. SHRaged lost more lactate dehydrogenase and tended to lose more xanthine and uric acid than other groups. CONCLUSIONS: The aged hypertrophied heart shows a higher vulnerability to ischaemic damage than the adult hypertrophied heart. This phenomenon is associated with subendocardial underperfusion, increased membrane damage and inadequate recovery of creatine phosphate levels.
Assuntos
Envelhecimento/fisiologia , Cardiomegalia/fisiopatologia , Coração/fisiopatologia , Hipertensão/fisiopatologia , Isquemia Miocárdica/fisiopatologia , Difosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Cardiomegalia/metabolismo , Hipertensão/metabolismo , Isquemia Miocárdica/metabolismo , Miocárdio/metabolismo , Perfusão , Fosfocreatina/metabolismo , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKYRESUMO
Knowledge of the effects of cardiopulmonary bypass on the myocardium and on cardiac function is limited. We therefore studied changes in haemodynamics and myocardial metabolism during the initial phase of cardiopulmonary bypass in two patient groups. In one group "normothermia" (34 degrees C) was used while on bypass, with an empty beating heart; in the other group hypothermia (range 27-33 degrees C) with ventricular fibrillation was used. Mean aortic pressure and myocardial oxygen consumption decreased significantly in both groups after instalment of CPB. The arterial-coronary sinus differences in lactate changed to negative values within 5 min of the start of bypass, indicating release instead of uptake of lactate. This release was maintained during the observation period and increased significantly in the hypothermic patient group when the ventricles were fibrillating. Therefore in patients undergoing aorto-coronary bypass surgery, detrimental changes in the myocardium must be anticipated during the initial phase of cardiopulmonary bypass prior to aortic cross clamping.
Assuntos
Aorta/fisiologia , Pressão Sanguínea , Ponte Cardiopulmonar , Ponte de Artéria Coronária , Miocárdio/metabolismo , Temperatura Corporal , Feminino , Hemodinâmica , Humanos , Lactatos/metabolismo , Masculino , Pessoa de Meia-Idade , Consumo de OxigênioRESUMO
A significant release of lactate instead of uptake was observed during the first 10 min of cardiopulmonary bypass preceding aorto-coronary bypass surgery in human patients. To clarify these findings in more detail, myocardial lactate and oxygen metabolism was studied in healthy dog hearts subjected to a protocol similar to the clinical situation. In one group (n = 11) normothermia at 34 degrees C was used with an empty beating heart, and in the other group (n = 11) hypothermia with ventricular fibrillation was applied. Within the first 10 min of bypass no significant changes in high energy phosphates were observed in myocardial biopsies. However, a marked decrease in mean aortic blood pressure and a simultaneous lowering in oxygen consumption was observed in both groups after instalment of bypass. An initial shift from lactate uptake to lactate release occurred while on bypass in the normothermia group. After 10 min of bypass, lactate uptake was restored in hearts of both groups. Therefore, the lactate release during the initial phase of bypass in patients originates both from the instalment of the bypass and from (local) inadequate perfusion, which is most likely to be due to stenosed coronary arteries.
Assuntos
Aorta/fisiologia , Pressão Sanguínea , Ponte Cardiopulmonar , Ponte de Artéria Coronária , Miocárdio/metabolismo , Animais , Temperatura Corporal , Cães , Hemodinâmica , Lactatos/metabolismo , Consumo de OxigênioRESUMO
OBJECTIVE: Previous studies have shown that chronic cardiac denervation impairs myocardial glucose oxidation. To investigate this further we tested whether the tissue content of glucose transporters, activity of glycolytic enzymes or metabolic capacity of pyruvate dehydrogenase were altered. Moreover, we investigated whether the decline in glucose utilization was associated with an upregulation of proteins and enzymes involved in fatty acid handling. Chronic cardiac denervation results also in decreased left ventricular efficiency. We explored whether alterations in mitochondrial properties could be held responsible for this phenomenon. METHODS: Twelve adult dogs were included in the study. In 6 of them chronic cardiac denervation was accomplished by surgical ablation of the extrinsic nerve fibers. The other 6 dogs were sham-operated. Biopsies were obtained from the left ventricle after 4-5 weeks of denervation. The content or enzymatic activity of proteins involved in fatty acid and glucose handling was assessed. Features of glutamate oxidation were measured in freshly isolated mitochondria. RESULTS: The content or activity of a set of fatty acid handling proteins did not change during chronic cardiac denervation. In contrast GLUT1 content significantly increased in the chronically denervated left ventricle, while the active form of pyruvate dehydrogenase declined (p < 0.05). Glutamate oxidation characteristics in freshly isolated mitochondria were not affected by chronic denervation. CONCLUSION: The impairment of glucose oxidation in the chronically denervated myocardium is most likely caused by a decline of pyruvate dehydrogenase in its active form. It is unlikely that the decrease in work efficiency is caused by alterations in mitochondrial properties.