RESUMO
Impaired platelet responsiveness to nitric oxide (NO resistance) is a common characteristic of many cardiovascular disease states and represents an independent risk factor for cardiac events and mortality. NO resistance reflects both scavenging of NO by superoxide (O2(-)), and impairment of the NO receptor, soluble guanylate cyclase (sGC). There is thus an urgent need for circumvention of NO resistance in order to improve clinical outcomes. Nitroxyl (HNO), like NO, produces vasodilator and anti-aggregatory effects, largely via sGC activation, but is not inactivated by O2(-). We tested the hypothesis that HNO circumvents NO resistance in human platelets. In 57 subjects with or without ischemic heart disease, platelet responses to the HNO donor isopropylamine NONOate (IPA/NO) and the NO donor sodium nitroprusside (SNP) were compared. While SNP (10µM) induced 29±3% (p<0.001) inhibition of platelet aggregation, IPA/NO (10µM) caused 75±4% inhibition (p<0.001). In NO-resistant subjects (n=28), the IPA/NO:SNP response ratio was markedly increased (p<0.01), consistent with partial circumvention of NO resistance. Similarly, cGMP accumulation in platelets was greater (p<0.001) with IPA/NO than with SNP stimulation. The NO scavenger carboxy-PTIO (CPTIO, 200µM) inhibited SNP and IPA/NO responses by 92±7% and 17±4% respectively (p<0.001 for differential inhibition), suggesting that effects of IPA/NO are only partially NO-mediated. ODQ (10µM) inhibited IPA/NO responses by 36±8% (p<0.001), consistent with a contribution of sGC/haem to IPA/NO inhibition of aggregation. There was no significant relationship between whole blood ROS content and IPA/NO responses. Thus the HNO donor IPA/NO substantially circumvents platelet NO resistance while acting, at least partially, as a haem-mediated sGC activator.
Assuntos
Plaquetas/efeitos dos fármacos , Óxido Nítrico/metabolismo , Óxidos de Nitrogênio/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Idoso , Idoso de 80 Anos ou mais , Plaquetas/metabolismo , Estudos de Casos e Controles , GMP Cíclico/metabolismo , Feminino , Humanos , Hidrazinas/farmacologia , Masculino , Pessoa de Meia-Idade , Isquemia Miocárdica , Nitroprussiato/farmacologia , OxirreduçãoRESUMO
AIMS/HYPOTHESIS: An increase in the production of reactive oxygen species is commonly thought to contribute to the development of diabetic cardiomyopathy. This study aimed to assess whether administration of the antioxidant coenzyme Q(10) would protect the diabetic heart against dysfunction and remodelling, using the db/db mouse model of type 2 diabetes. Furthermore, we aimed to compare the efficacy of coenzyme Q(10) to that of the ACE inhibitor ramipril. METHODS: Six-week-old non-diabetic db/+ mice and diabetic db/db mice received either normal drinking water or water supplemented with coenzyme Q(10) for 10 weeks. Endpoint cardiac function was assessed by echocardiography and catheterisation. Ventricular tissue was collected for histology, gene expression and protein analysis. RESULTS: Untreated db/db diabetic mice exhibited hyperglycaemia, accompanied by diastolic dysfunction and adverse structural remodelling, including cardiomyocyte hypertrophy, myocardial fibrosis and increased apoptosis. Systemic lipid peroxidation and myocardial superoxide generation were also elevated in db/db mice. Coenzyme Q(10) and ramipril treatment reduced superoxide generation, ameliorated diastolic dysfunction and reduced cardiomyocyte hypertrophy and fibrosis in db/db mice. Phosphorylation of Akt, although depressed in untreated db/db mice, was restored with coenzyme Q(10) administration. We postulate that preservation of cardioprotective Akt signalling may be a mechanism by which coenzyme Q(10)-treated db/db mice are protected from pathological cardiac hypertrophy. CONCLUSIONS/INTERPRETATION: These data demonstrate that coenzyme Q(10) attenuates oxidative stress and left ventricular diastolic dysfunction and remodelling in the diabetic heart. Addition of coenzyme Q(10) to the current therapy used in diabetic patients with diastolic dysfunction warrants further investigation.
Assuntos
Cardiomegalia/tratamento farmacológico , Cardiomiopatias Diabéticas/tratamento farmacológico , Ubiquinona/análogos & derivados , Vitaminas/uso terapêutico , Animais , Anti-Hipertensivos/uso terapêutico , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Cardiomegalia/diagnóstico por imagem , Cardiomegalia/metabolismo , Cardiomegalia/fisiopatologia , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatologia , Cardiomiopatias Diabéticas/diagnóstico por imagem , Cardiomiopatias Diabéticas/metabolismo , Cardiomiopatias Diabéticas/fisiopatologia , Modelos Animais de Doenças , Fibrose Endomiocárdica/tratamento farmacológico , Fibrose Endomiocárdica/etiologia , Fibrose Endomiocárdica/metabolismo , Feminino , Hiperglicemia/tratamento farmacológico , Hiperglicemia/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ramipril/uso terapêutico , Superóxidos/antagonistas & inibidores , Superóxidos/metabolismo , Ubiquinona/uso terapêutico , Ultrassonografia , Remodelação Ventricular/efeitos dos fármacos , Remodelação Ventricular/fisiologiaRESUMO
AIMS/HYPOTHESIS: Diabetic cardiomyopathy is characterised by diastolic dysfunction, oxidative stress, fibrosis, apoptosis and pathological cardiomyocyte hypertrophy. Phosphoinositide 3-kinase (PI3K)(p110α) is a cardioprotective kinase, but its role in the diabetic heart is unknown. The aim of this study was to assess whether PI3K(p110α) plays a critical role in the induction of diabetic cardiomyopathy, and whether increasing PI3K(p110α) activity in the heart can prevent the development of cardiac dysfunction in a setting of diabetes. METHODS: Type 1 diabetes was induced with streptozotocin in adult male cardiac-specific transgenic mice with increased PI3K(p110α) activity (constitutively active PI3K [p110α], caPI3K] or decreased PI3K(p110α) activity (dominant-negative PI3K [p110α], dnPI3K) and non-transgenic (Ntg) mice for 12 weeks. Cardiac function, histological and molecular analyses were performed. RESULTS: Diabetic Ntg mice displayed diastolic dysfunction and increased cardiomyocyte size, expression of atrial and B-type natriuretic peptides (Anp, Bnp), fibrosis and apoptosis, as well as increased superoxide generation and increased protein kinase C ß2 (PKCß2), p22 ( phox ) and apoptosis signal-regulating kinase 1 (Ask1) expression. Diabetic dnPI3K mice displayed an exaggerated cardiomyopathy phenotype compared with diabetic Ntg mice. In contrast, diabetic caPI3K mice were protected against diastolic dysfunction, pathological cardiomyocyte hypertrophy, fibrosis and apoptosis. Protection in diabetic caPI3K mice was associated with attenuation of left ventricular superoxide generation, attenuated Anp, Bnp, PKCß2, Ask1 and p22 ( phox ) expression, and elevated AKT. Further, in cardiomyocyte-like cells, increased PI3K(p110α) activity suppressed high glucose-induced superoxide generation and enhanced mitochondrial function. CONCLUSIONS/INTERPRETATION: These results demonstrate that reduced PI3K activity accelerates the development of diabetic cardiomyopathy, and that enhanced PI3K(p110α) activity can prevent adverse cardiac remodelling and dysfunction in a setting of diabetes.
Assuntos
Classe I de Fosfatidilinositol 3-Quinases/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Cardiomiopatias Diabéticas/metabolismo , Cardiomiopatias Diabéticas/prevenção & controle , Superóxidos/metabolismo , Animais , Northern Blotting , Regulação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Estresse OxidativoRESUMO
BACKGROUND: Nitric oxide (NO) is a modulator of left ventricular hypertrophy (LVH) and myocardial relaxation. The impact of NO availability on development of LVH has never been demonstrated in humans. We tested the hypotheses that elevation of asymmetric dimethylarginine (ADMA) concentrations (biochemical marker of decreased NO generation), and impairment of vascular responsiveness to NO donor GTN, would each predict the presence of LVH and associated LV diastolic dysfunction in a normal aging population. METHODS AND RESULTS: In 74 subjects aged 68±6 years, LV volumes and mass indexed to height(2.7) (LVMI) were calculated from cardiac MRI. Despite the absence of clinically-defined LVH, there was a relationship (r=0.29; p=0.01) between systolic BP and LVMI. Both elevation of ADMA levels to the highest quartile or impairment of GTN responsiveness (determined by applanation tonometry) to the lowest quartile were determinants of LVMI independent of systolic BP (p=0.01 and p=0.03, respectively). Filling pressure (E/E' ratio from echocardiography) was increased in patients with impaired vascular NO responsiveness (p<0.05) irrespective of LVMI. ADMA remained a significant determinant of LVMI on multivariate analysis. CONCLUSIONS: These data imply that NO bioavailability within the myocardium modulates earliest stages of LVH development and facilitates development of diastolic dysfunction at a given LV mass.
Assuntos
Arginina/análogos & derivados , Hipertrofia Ventricular Esquerda/sangue , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico/metabolismo , Idoso , Idoso de 80 Anos ou mais , Arginina/sangue , Arginina/metabolismo , Pressão Sanguínea , Ecocardiografia , Feminino , Humanos , Hipertrofia Ventricular Esquerda/diagnóstico , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Miocárdio/metabolismo , Óxido Nítrico/análise , Óxido Nítrico Sintase/sangue , Valor Preditivo dos Testes , SoftwareRESUMO
Monte Carlo computer models that simulate the detailed, event-by-event transport of electrons in liquid water are valuable for the interpretation and understanding of findings in radiation chemistry and radiation biology. Because of the paucity of experimental data, such efforts must rely on theoretical principles and considerable judgment in their development. Experimental verification of numerical input is possible to only a limited extent. Indirect support for model validity can be gained from a comparison of details between two independently developed computer codes as well as the observable results calculated with them. In this study, we compare the transport properties of electrons in liquid water using two such models, PARTRAC and NOREC. Both use interaction cross sections based on plane-wave Born approximations and a numerical parameterization of the complex dielectric response function for the liquid. The models are described and compared, and their similarities and differences are highlighted. Recent developments in the field are discussed and taken into account. The calculated stopping powers, W values, and slab penetration characteristics are in good agreement with one another and with other independent sources.
Assuntos
Elétrons , Modelos Químicos , Água/química , Simulação por Computador , Íons/química , Método de Monte Carlo , Reprodutibilidade dos TestesRESUMO
Angiotensin-converting enzyme inhibitors block left ventricular hypertrophy in vivo. A component of this effect has been attributed to tissue accumulation of bradykinin. Little is known regarding the effect of bradykinin on cardiomyocytes. The objectives of the present study were to define the effects of bradykinin on isolated ventricular cardiomyocytes (from adult and neonatal rat hearts) and to determine the extent to which bradykinin blocks hypertrophy in vitro. Bradykinin was found to be a hypertrophic agonist, as defined by increased protein synthesis and atrial natriuretic peptide secretion and expression. Bradykinin (10 micromol/L) increased [3H]phenylalanine incorporation by 23+/-3% in adult and by 36+/-10% in neonatal cardiomyocytes. Constitutive atrial natriuretic peptide secretion by neonatal myocytes was increased 357+/-103%. All effects of bradykinin were abolished by the B2-kinin receptor antagonist Hoe 140. These increases were similar in magnitude to those observed with phenylephrine (20 micromol/L) and angiotensin II (1 micromol/L). However, in cardiomyocytes cocultured with endothelial cells, bradykinin did not increase protein synthesis. Angiotensin II increased [3H]phenylalanine incorporation by 24+/-3% in adult cardiomyocytes in monoculture and by 22+/-2% in adult rat cardiomyocytes cocultured with endothelial cells. Bradykinin abolished this angiotensin II-induced hypertrophy in myocytes cultured with endothelial cells but not in myocytes studied in the absence of endothelial cells. In conclusion, bradykinin has a direct hypertrophic effect on ventricular myocytes. The presence of endothelial cells is required for the antihypertrophic effects of bradykinin. The results suggest that the increase in local concentration of bradykinin associated with angiotensin-converting enzyme inhibition is an important mechanism by which hypertrophy can be blocked. Manifestation of this mechanism appears to require bradykinin-stimulated release of paracrine factor(s) from endothelial cells, which are also able to block the hypertrophic effects of Ang II.
Assuntos
Bradicinina/farmacologia , Hipertrofia Ventricular Esquerda/induzido quimicamente , Angiotensina II/farmacologia , Animais , Animais Recém-Nascidos , Bradicinina/análogos & derivados , Antagonistas dos Receptores da Bradicinina , Células Cultivadas , Endotélio/citologia , Ventrículos do Coração/citologia , Ventrículos do Coração/efeitos dos fármacos , Hipertrofia Ventricular Esquerda/prevenção & controle , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
The spatial distributions of ionizations and other inelastic events in charged-particle tracks are important quantities that influence the final outcome of radiation interaction. Calculations of such distributions are presented for the tracks of electrons in the energy range 100 eV to 10 keV in liquid water and water vapor, and the results are compared. The distributions include the frequency of nearest-neighbor distances for all inelastic events, the mean nearest-neighbor distances for ionizations and for all inelastic events as a function of electron energy, the frequency of distances between all ionizations and all inelastic events, and the farthest distances between all inelastic events in electron tracks. The physical differences between liquid water and water vapor are discussed in terms of the respective inverse mean free paths, the collision spectra, and the nonlocalization of energy losses that are likely to occur in the liquid.
Assuntos
Elétrons , Soluções , Volatilização , ÁguaRESUMO
Detailed Monte Carlo calculations have been carried out of slowing-down spectra and yields for a number of end-points for electrons in liquid water. These investigations were made to study differences in physical effects of different low-LET radiations and implications for RBE. Initial electron energies from 1 keV to 1 MeV were used, and all secondary electrons were followed in the computations unitl their energies fell below 10 eV. Though there are substantial differences in the slowing-down spectra at energies near and above the K-shell ionisation potential of oxygen, the energy spectrum of electrons at lower energies is found to be essentially independent of the initial energy of the primary electron. The number of events per unit energy deposited is also essentially independent of the primary electron energy. Based on these calculations, there appears to be little basis for ascribing differences in RBE for low-LET radiations to differences in physical effects produced by secondary electrons of low energy (less than or equal to 1 keV).
Assuntos
Elétrons , Transferência de Energia , Método de Monte Carlo , Radiometria/instrumentação , ÁguaRESUMO
We have recently reported the time course of acute myocardial drug uptake and simultaneous pharmacodynamic effects for metoprolol (4 mg; n = 12) and sotalol (20 mg; n = 10) in patients with ischemic heart disease. The acute pharmacodynamic effects of the two drugs included reductions in both spontaneous heart rate and the contractile index peak positive rate of left ventricular pressure rise and prolongation of the electrocardiographic PR interval, all of which exhibited an equilibration delay compared with myocardial drug content. The objective of the current study was to analyze the relationship between myocardial drug content and effect for both metoprolol and sotalol using an effect compartment model, to examine the potential for the two drugs to share a common subsite of action in the myocardium. The time course of myocardial drug content was best described by a one-compartment model for metoprolol and a two-compartment model for sotalol. A linear pharmacodynamic model, relating the amount of drug at the effect-site (Ae) with each of the three effects, was used in the effect compartment modeling. The slope of each of these effects as a function of Ae was considerably flatter for sotalol than for metoprolol, reflecting the relative beta-adrenoceptor antagonistic potencies of the two drugs. The exit rate constant from the effect compartment (K(eo)) did not differ significantly from each other or from the exit rate constant from the "peripheral" compartment (K21) for sotalol. The results suggest that the effect compartment within the myocardium for both drugs may correspond to a "peripheral" compartment, even though a "peripheral" pharmacokinetic compartment for myocardial metoprolol content was not apparent. Thus, the effect compartment for many cardioactive drugs may correspond to a pharmacokinetically "peripheral" compartment, irrespective of localization of effect to a small (e.g. sinoatrial node) or large (e.g. ventricular myocardium) region of the heart.
Assuntos
Antagonistas Adrenérgicos beta/farmacocinética , Antiarrítmicos/farmacocinética , Metoprolol/farmacocinética , Miocárdio/metabolismo , Sotalol/farmacocinética , Antagonistas Adrenérgicos beta/farmacologia , Antiarrítmicos/farmacologia , Área Sob a Curva , Depressão Química , Frequência Cardíaca/efeitos dos fármacos , Humanos , Injeções Intravenosas , Modelos Lineares , Metoprolol/farmacologia , Modelos Biológicos , Contração Miocárdica/efeitos dos fármacos , Miocárdio/química , Análise de Regressão , Sotalol/farmacologiaRESUMO
BACKGROUND AND PURPOSE: While maintaining cardiac performance, chronic ß-adrenoceptor activation eventually exacerbates the progression of cardiac remodelling and failure. We examined the adverse signalling pathways mediated by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and reactive oxygen species (ROS) after chronic ß2-adrenoceptor activation. EXPERIMENTAL APPROACH: Mice with transgenic ß2-adrenoceptor overexpression (ß2-TG) and non-transgenic littermates were either untreated or treated with an antioxidant (N-acetylcysteine, NAC) or NADPH oxidase inhibitors (apocynin, diphenyliodonium). Levels of ROS, phosphorylated p38 mitogen-activated protein kinase (MAPK), pro-inflammatory cytokines and collagen content in the left ventricle (LV) and LV function were measured and compared. KEY RESULTS: ß2-TG mice showed increased ROS production, phosphorylation of p38 MAPK and heat shock protein 27 (HSP27), expression of pro-inflammatory cytokines and collagen, and progressive ventricular dysfunction. ß2-adrenoceptor stimulation similarly increased ROS production and phosphorylation of p38 MAPK and HSP27 in cultured cardiomyocytes. Treatment with apocynin, diphenyliodonium or NAC reduced phosphorylation of p38 MAPK and HSP27 in both cultured cardiomyocytes and the LV of ß2-TG mice. NAC treatment (500 mg·kg⻹ ·day⻹) for 2 weeks eliminated the up-regulated expression of pro-inflammatory cytokines and collagen in the LV of ß2-TG mice. Chronic NAC treatment to ß2-TG mice from 7 to 10 months of age largely prevented progression of ventricular dilatation, preserved contractile function (fractional shortening 37 ± 5% vs. 25 ± 3%, ejection fraction 52 ± 5% vs. 32 ± 4%, both P < 0.05), reduced cardiac fibrosis and suppressed matrix metalloproteinase activity. CONCLUSION AND IMPLICATIONS: ß2-adrenoceptor stimulation provoked NADPH oxidase-derived ROS production in the heart. Elevated ROS activated p38 MAPK and contributed significantly to cardiac inflammation, remodelling and failure.
Assuntos
Cardiomiopatias/etiologia , Cardiomiopatias/metabolismo , Insuficiência Cardíaca/etiologia , Insuficiência Cardíaca/metabolismo , Miocárdio/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Acetilcisteína/farmacologia , Animais , Antioxidantes/farmacologia , Cardiomiopatias/tratamento farmacológico , Colágeno/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Proteínas de Choque Térmico HSP27/metabolismo , Insuficiência Cardíaca/tratamento farmacológico , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Miócitos Cardíacos/metabolismo , NADPH Oxidases/antagonistas & inibidores , NADPH Oxidases/metabolismo , Estresse Oxidativo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Espécies Reativas de Oxigênio/metabolismo , Receptores Adrenérgicos beta 2/genética , Transdução de Sinais , Remodelação Ventricular/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoAssuntos
Alumínio/análise , Transferência de Energia , Cobre , Elétrons , Isótopos de Ouro , Radioisótopos , Análise EspectralRESUMO
Reactive oxygen species such as superoxide are implicated in cardiac hypertrophy, but their contribution to the cardiac complications of insulin resistance is unresolved. We tested the hypothesis that the antioxidant tempol attenuates cardiac hypertrophy in insulin-resistant mice. Mice with cardiac GLUT4 deletion (GLUT4-knockout), superimposed on global GLUT4 suppression (GLUT4-knockdown) were administered tempol for 4 weeks. Age-matched GLUT4-knockdown littermates were used as controls (14 mice/group). GLUT4-knockout mice exhibited marked cardiac hypertrophy: heart to body weight ratio was increased 61+/-7% and expression of the hypertrophic genes beta-myosin heavy chain and B-type natriuretic peptide (BNP) were elevated 5.5+/-0.7- and 6.2+/-1.5-fold relative to control, respectively. Pro-fibrotic pro-collagen III expression was also higher (3.8+/-0.7-fold) in the GLUT4-knockout myocardium (all p<0.001). Both gp91(phox) and Nox1 subunits of NADPH oxidase were also upregulated, 4.9+/-1.2- and 9.3+/-2.8-fold (both p<0.01). Tempol treatment significantly attenuated all of these abnormalities in GLUT4-knockout mice. Heart to body weight ratio was decreased, as was fold expression of beta-myosin heavy chain (to 3.8+/-0.8), BNP (to 2.5+/-0.5), pro-collagen III (to 1.9+/-0.4), gp91(phox) (to 0.9+/-0.3) and Nox1 (to 2.3+/-0.1, all p<0.05 versus untreated GLUT4-knockout mice). In addition, tempol upregulated ventricular expression of both thioredoxin-2 (confirming an antioxidant action) and glycogen synthase kinase-3beta (GSK-3beta). Tempol did not elicit any other significant changes in control mice. Cardiac superoxide generation, however, was not altered by GLUT4-knockout or tempol. In conclusion, tempol treatment reduced morphological and molecular evidence of cardiac hypertrophy in the GLUT4-knockout insulin-resistant mouse in vivo, even at doses insufficient to lower cardiac superoxide. Parallel reductions in pro-collagen III and NADPH oxidase have important implications for our understanding of the molecular basis of cardiac hypertrophy in the setting of insulin resistance. Antioxidants may offer new alternatives in this disorder.
Assuntos
Antioxidantes/farmacologia , Cardiomegalia/tratamento farmacológico , Óxidos N-Cíclicos/farmacologia , Transportador de Glucose Tipo 4/deficiência , Resistência à Insulina/genética , Animais , Feminino , Transportador de Glucose Tipo 4/genética , Masculino , Camundongos , Camundongos Knockout , Marcadores de SpinRESUMO
1. The influence of hypoxia on the time course of regional myocardial accumulation and elimination of the beta 1-adrenoceptor antagonist metoprolol was investigated by a spontaneously beating rat isolated perfused heart preparation. 2. Myocardial metoprolol content was maximal at 2 min in the left and right ventricles and atria. Neither the extent nor the time of maximal myocardial metoprolol content was significantly influenced by the induction of hypoxia. However, maximal myocardial metoprolol content in both atria and right ventricles was significantly higher than that in the left ventricular samples (P < 0.02; one-factor analysis of variance, 17 d.f.). 3. Elimination of metoprolol (as indicated from residual myocardial metoprolol content at 10 min) was impaired in hypoxic left ventricles (P < 0.01 vs normoxia; unpaired t-test, 10 d.f.) but not in right ventricles or atria. This variation in myocardial metoprolol disposition was not apparent from examination of serial metoprolol concentrations in coronary perfusate. 4. Hypoxia selectively impaired the elimination of metoprolol from the left ventricle, but not the process of drug accumulation, by any region of myocardium. It remains to be determined whether this reflects regional variation in the extent of microcirculatory impairment associated with hypoxia.
Assuntos
Antagonistas Adrenérgicos beta/farmacocinética , Hipóxia/metabolismo , Metoprolol/farmacocinética , Miocárdio/metabolismo , Antagonistas Adrenérgicos beta/sangue , Animais , Átrios do Coração/metabolismo , Ventrículos do Coração/metabolismo , Técnicas In Vitro , Masculino , Metoprolol/sangue , Perfusão , Ratos , Ratos Sprague-DawleyRESUMO
Most of the energy absorbed in the cell nucleus from a radiation field goes into the aqueous medium that surrounds macromolecules, like DNA, which are critical to the normal function of cells. This part of the energy deposition produces numerous reactive species that can diffuse to DNA sequences and induce chemical changes. The average diffusion distance of the free radicals that mediate this indirect mode of DNA damage is only a few nanometers because the cellular medium contains a high concentration of molecules that rapidly scavenge the radiation-induced species. Under these conditions, direct interaction of the radiation field with the DNA can not be neglected as a potential mode of damage induction. Two aspects of the direct effect will be discussed in this paper: (1) screening of the interaction between DNA and charged particles by the dielectric response of the aqueous medium and (2) the impact-parameter dependence of these interactions.
Assuntos
Dano ao DNA , DNA/química , Conformação de Ácido Nucleico , Núcleo Celular/metabolismo , DNA/metabolismo , DNA/efeitos da radiação , Transferência de Energia , Íons , Matemática , Modelos Estruturais , Modelos Teóricos , Fótons , SoluçõesRESUMO
We have previously demonstrated that bradykinin blocks hypertrophy of isolated cardiomyocytes: this is dependent on the release of nitric oxide from endothelial cells. In the present study, we investigated the influence of endothelial dysfunction on the antihypertrophic action of bradykinin. Angiotensin II (1 microM) induced a 34 +/- 2% increase in [3H]phenylalanine incorporation (P<0.001), an in vitro marker of hypertrophy, in adult rat cardiomyocytes co-cultured with bovine aortic endothelial cells. This response was blocked by bradykinin (10 microM), but restored by the nitric oxide synthase inhibitor. N(omega)-monomethyl-L-arginine (100 microM). However, the antihypertrophic effect of bradykinin in co-culture was abolished by 24 h pretreatment of endothelial cells with high glucose (25 mM, to mimic hyperglycemia) and attenuated by hydrogen peroxide (100 microM, to mimic oxidative stress). Pretreatment with oxidized low-density lipoprotein (100 microg/ml for 24 h, to mimic hyperlipidemia) was without effect. The hypertrophic response to angiotensin II was not modified by endothelial cell pretreatment. Furthermore, the ability of bradykinin to elevate cGMP (a marker for nitric oxide) in cardiomyocytes co-cultured with endothelial cells was attenuated by pretreatment with either high glucose or hydrogen peroxide. In conclusion, loss of the cardioprotective action of bradykinin against angiotensin II-induced hypertrophy was associated with impaired nitric oxide release from dysfunctional endothelial cells.
Assuntos
Bradicinina/farmacologia , Endotélio Vascular/fisiopatologia , Coração/efeitos dos fármacos , Angiotensina II/metabolismo , Angiotensina II/farmacologia , Animais , Bradicinina/metabolismo , Bovinos , Células Cultivadas , GMP Cíclico/metabolismo , Masculino , Miocárdio/citologia , Ratos , Ratos Sprague-DawleyRESUMO
1. The extent of postextrasystolic potentiation (PESP) has been considered a useful parameter for evaluating myocardial contractile reserve in the presence of myocardial stunning or hibernation. Extent of PESP appears to reflect an interaction between myofibrillar calcium concentration and function of the contractile apparatus. However, potential for cardiovascular drugs including agents modifying adenosine 3' 5'-cyclic monophosphate concentration to influence the extent of PESP in man has not been extensively studied. 2. In 35 patients undergoing diagnostic coronary angiography, we investigated the relationship between the extrasystolic test pulse interval (ETPI) and left ventricular (LV) +dP/dtmax of a postextrasystolic contraction. The influence of three inotropically active agents on this relationship was examined following intravenous bolus injection (metoprolol, 4 mg; sotalol, 20 mg; and milrinone, 1 mg). 3. The patient group examined had predominantly preserved LV function (LVEF 67% with 95% confidence intervals 63%, 71%). In the doses utilized, all agents exerted significant effects on LV+dP/dtmax during atrial pacing: reduction of 12.3% (6.4, 18.2) for metoprolol (P < 0.0005), and 10.9% (4.2, 17.6) for sotalol (P < 0.005); and increase of 11.8% (1.3, 22.3) for milrinone (P < 0.05). 4. With the postextrasystolic interval identical to baseline pacing cycle length, postextrasystolic potentiation of LV+dP/dtmax varied inversely with ETPI. None of the three agents investigated significantly affected this relationship. 5. These results demonstrate that the extent of PESP is unaffected by 'pure' beta-adrenoceptor antagonism, (+/-)-sotalol or phosphodiesterase inhibition in man. Hence pharmacotherapy with these agents is unlikely to affect assessment of extent of PESP.
Assuntos
Antagonistas Adrenérgicos beta/farmacologia , Complexos Cardíacos Prematuros/fisiopatologia , Cardiotônicos/farmacologia , Contração Miocárdica/efeitos dos fármacos , Isquemia Miocárdica/fisiopatologia , Função Ventricular Esquerda/efeitos dos fármacos , Adulto , Idoso , Complexos Cardíacos Prematuros/tratamento farmacológico , Feminino , Humanos , Injeções Intravenosas , Masculino , Metoprolol/farmacologia , Pessoa de Meia-Idade , Milrinona , Isquemia Miocárdica/tratamento farmacológico , Miocárdio Atordoado/fisiopatologia , Inibidores de Fosfodiesterase/farmacologia , Piridonas/farmacologia , Sotalol/farmacologia , Função Ventricular Esquerda/fisiologiaRESUMO
Bradykinin (BK) has a direct hypertrophic effect on rat ventricular cardiomyocytes (VCM) as defined by an increase in protein synthesis and an increase in atrial natriuretic peptide mRNA and secretion. In the current study, we have examined the dependence of BK-induced protein synthesis on activation of 90-kDa ribosomal S6 kinase (p90(rsk)) and 70-kDa S6 kinase (p70(S6K)). Both of these kinases possess the ability to phosphorylate the ribosomal protein S6, which plays an important role in initiating mRNA translation. Stimulation of adult VCM with 10 microM BK increased p90(rsk) activity by 2.5 +/- 0.3-fold and increased p70(S6K) activity by 2.0 +/- 0.3-fold. p90(rsk) is a terminal kinase in the mitogen-activated protein (MAP) kinase pathway. Inhibition of MAP kinase kinase activation by Raf in the MAP kinase pathway with PD-098059 (25 microM) blocked BK-stimulated activation of p90(rsk) by 70% and unexpectedly blocked p70(S6K) by 72%. Rapamycin inhibited BK-stimulated p70(S6K) activity by 93% but had no effect on p90(rsk) activation by BK. Inhibition of the MAP kinase pathway and p70(S6K) with PD-098059 was paralleled by changes in protein synthesis. BK (10 microM) increased [3H]phenylalanine incorporation by 27 +/- 3 and 39 +/- 6% in cultured adult and neonatal VCM, respectively. Treatment with PD-098059 or rapamycin abolished the increase in protein synthesis stimulated by BK. These results suggest that 1) BK activates p70(S6K) and p90(rsk); 2) although both p70(S6K) and p90(rsk) have the potential to phosphorylate the ribosomal S6 protein, p70(S6K) and not p90(rsk) is the predominant kinase involved in increasing protein synthesis by BK; and 3) p70(S6K) activation is dependent on stimulation of the MAP kinase pathway at a point distal to Raf.
Assuntos
Bradicinina/farmacologia , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Proteínas Musculares/biossíntese , Miocárdio/metabolismo , Proteínas Quinases S6 Ribossômicas/fisiologia , Angiotensina II/farmacologia , Animais , Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Masculino , Proteínas Musculares/antagonistas & inibidores , Miocárdio/citologia , Fenilalanina/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Quinases S6 Ribossômicas/antagonistas & inibidores , Proteínas Quinases S6 Ribossômicas/metabolismo , Sirolimo/farmacologiaRESUMO
One of the major determinants of the short-term effects of many cardioactive drugs is the concentration of the drug specifically within the myocardium. However, no information regarding the disposition of the phosphodiesterase inhibitor milrinone in the heart is available. We therefore determined the time course of short-term myocardial milrinone uptake from paired transcoronary sampling and simultaneous coronary sinus blood flow after a 1-mg intravenous bolus in patients undergoing diagnostic cardiac catheterization. In accordance with this intention, a sensitive, reproducible method for the determination of milrinone in human whole-blood samples was developed. The reverse-phase high-performance liquid chromatographic method described used a C18 column with UV-absorbance detection at 326 nm, with a limit of detection of 0.6 ng/ml, and was highly reproducible. The short-term hemodynamic and electrophysiologic effects of the drug also were determined. Significant increases in spontaneous heart rate and LV+dP/dtmax (at constant heart rate) were observed, accompanied by reductions in mean arterial pressure, systemic vascular resistance, and PR interval, without significant changes in atrioventricular nodal or ventricular effective refractory periods. Peak content (1.89 +/- 0.30% of injected dose) was rapidly attained, 0.56 +/- 0.06 min after milrinone injection. Time of peak effects was significantly delayed (7-10 min after injection) relative to time of peak myocardial milrinone content. Residual myocardial milrinone content was 69.1 +/- 5.7% of maximum 12.5 min after injection. It is concluded that both myocardial uptake and the onset of positive inotropic effects after intravenous injection of milrinone were very rapid. However, there was significant hysteresis between peak myocardial content and subsequent hemodynamic effects.