RESUMEN
We cloned and sequenced two cDNAs encoding the angiogenic, vascular endothelial growth factor (VEGF) from the porcine heart. Deduced amino acid sequence of the clone pPVE-18 and pPVE-5 predicted 164 (VEGF164), and 120 (VEGF120) residues of VEGF, respectively, with a putative N-terminal signal sequence of 26 amino acids. The porcine VEGF is shorter by one amino acid as compared to human VEGF, but a potential glycosylation site is present at Asn-74. PCR detection, and verification of the identity of the PCR products by Southern hybridization, confirmed wide expression of VEGF in different porcine tissues. Northern blot analysis with a radiolabeled porcine specific VEGF probe, showed one major (3.9 kb) and one minor (1.7 kb) mRNA species expressed in all four chambers of the heart.
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Endotelio Vascular/metabolismo , Sustancias de Crecimiento/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario/biosíntesis , Expresión Génica , Sustancias de Crecimiento/química , Datos de Secuencia Molecular , Alineación de Secuencia , PorcinosRESUMEN
BACKGROUND: Elevated concentrations of norepinephrine (NE) have been observed in ischemic myocardium. We investigated the magnitude and mechanism of catecholamine release in the myocardial interstitial fluid (MIF) during ischemia and reperfusion in vivo through the use of microdialysis. METHODS AND RESULTS: In 9 anesthetized pigs, interstitial catecholamine concentrations were measured in the perfusion areas of the left anterior descending coronary artery (LAD) and the left circumflex coronary artery. After stabilization, the LAD was occluded for 60 minutes and reperfused for 150 minutes. During the final 30 minutes, tyramine (154 nmol. kg(-1). min(-1)) was infused into the LAD. During LAD occlusion, MIF NE concentrations in the ischemic region increased progressively from 1. 0+/-0.1 to 524+/-125 nmol/L. MIF concentrations of dopamine and epinephrine rose from 0.4+/-0.1 to 43.9+/-9.5 nmol/L and from <0.2 (detection limit) to 4.7+/-0.7 nmol/L, respectively. Local uptake-1 blockade attenuated release of all 3 catecholamines by >50%. During reperfusion, MIF catecholamine concentrations returned to baseline within 120 minutes. At that time, the tyramine-induced NE release was similar to that seen in nonischemic control animals despite massive infarction. Arterial and MIF catecholamine concentrations in the left circumflex coronary artery region remained unchanged. CONCLUSIONS: Myocardial ischemia is associated with a pronounced increase of MIF catecholamines, which is at least in part mediated by a reversed neuronal reuptake mechanism. The increase of MIF epinephrine implies a (probably neuronal) cardiac source, whereas the preserved catecholamine response to tyramine in postischemic necrotic myocardium indicates functional integrity of sympathetic nerve terminals.
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Infarto del Miocardio/metabolismo , Isquemia Miocárdica/metabolismo , Norepinefrina/metabolismo , Sistema Nervioso Simpático/metabolismo , Animales , Presión Sanguínea/fisiología , Circulación Coronaria/fisiología , Femenino , Corazón/inervación , Corazón/fisiología , Frecuencia Cardíaca/fisiología , Masculino , Microdiálisis , Infarto del Miocardio/fisiopatología , Isquemia Miocárdica/fisiopatología , Reperfusión Miocárdica , Terminaciones Nerviosas/metabolismo , Volumen Sistólico/fisiología , Porcinos , Sistema Nervioso Simpático/efectos de los fármacos , Simpatomiméticos/farmacología , Tiramina/farmacología , Fibrilación Ventricular/metabolismoRESUMEN
BACKGROUND: The mechanisms behind the beneficial effects of renin-angiotensin system blockade after myocardial infarction (MI) are not fully elucidated but may include interference with tissue angiotensin II (Ang II). METHODS AND RESULTS: Forty-nine pigs underwent coronary artery ligation or sham operation and were studied up to 6 weeks. To determine coronary angiotensin I (Ang I) to Ang II conversion and to distinguish plasma-derived Ang II from locally synthesized Ang II, (125)I-labeled and endogenous Ang I and II were measured in plasma and in infarcted and noninfarcted left ventricle (LV) during (125)I-Ang I infusion. Ang II type 1 (AT(1)) receptor-mediated uptake of circulating (125)I-Ang II was increased at 1 and 3 weeks in noninfarcted LV, and this uptake was the main cause of the transient elevation in Ang II levels in the noninfarcted LV at 1 week. Ang II levels and AT(1) receptor-mediated uptake of circulating Ang II were reduced in the infarct area at all time points. Coronary Ang I to Ang II conversion was unaffected by MI. Captopril and the AT(1) receptor antagonist eprosartan attenuated postinfarct remodeling, although both drugs increased cardiac Ang II production. Captopril blocked coronary conversion by >80% and normalized Ang II uptake in the noninfarcted LV. Eprosartan did not affect coronary conversion and blocked cardiac Ang II uptake by >90%. CONCLUSIONS: Both circulating and locally generated Ang II contribute to remodeling after MI. The rise in tissue Ang II production during angiotensin-converting enzyme inhibition and AT(1) receptor blockade suggests that the antihypertrophic effects of these drugs result not only from diminished AT(1) receptor stimulation but also from increased stimulation of growth-inhibitory Ang II type 2 receptors.
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Modelos Cardiovasculares , Infarto del Miocardio/sangre , Peptidil-Dipeptidasa A/sangre , Receptores de Angiotensina/sangre , Angiotensina II/sangre , Animales , Captopril/sangre , Receptor de Angiotensina Tipo 1 , Receptor de Angiotensina Tipo 2 , PorcinosRESUMEN
BACKGROUND: Radioactive stents have been reported to reduce in-stent neointimal thickening. An unexpected increase in neointimal response was observed, however, at the stent-to-artery transitions, the so-called "edge effect." To investigate the factors involved in this edge effect, we studied stents with 1 radioactive half and 1 regular nonradioactive half, thereby creating a midstent radioactive dose-falloff zone next to a nonradioactive stent-artery transition at one side and a radioactive stent-artery transition at the other side. METHODS AND RESULTS: Half-radioactive stents (n=20) and nonradioactive control stents (n=10) were implanted in the coronary arteries of Yucatan micropigs. Animals received aspirin and clopidogrel as antithrombotics. After 4 weeks, a significant midstent stenosis was observed by angiography in the half-radioactive stents. Two animals died suddenly because of coronary occlusion at this mid zone at 8 and 10 weeks. At 12-week follow-up angiography, intravascular ultrasound and histomorphometry showed a significant neointimal thickening at the midstent dose-falloff zone of the half-radioactive stents, but not at the stent-to-artery transitions at both extremities. Such a midstent response (mean angiographic late loss 1.0 mm) was not observed in the nonradioactive stents (mean loss 0.4 to 0.6 mm; P< 0.01). CONCLUSIONS: The edge effect of high-dose radioactive stents in porcine coronary arteries is associated with the combination of stent injury and radioactive dose falloff.
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Vasos Coronarios/efectos de la radiación , Oclusión de Injerto Vascular/etiología , Oclusión de Injerto Vascular/prevención & control , Radioisótopos de Fósforo/administración & dosificación , Stents/efectos adversos , Animales , Implantación de Prótesis Vascular , Angiografía Coronaria , Vasos Coronarios/patología , Vasos Coronarios/cirugía , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Relación Dosis-Respuesta en la Radiación , Implantes de Medicamentos , Femenino , Oclusión de Injerto Vascular/patología , Implantes Experimentales , Porcinos Enanos , Túnica Íntima/patología , Túnica Íntima/efectos de la radiación , Grado de Desobstrucción Vascular/efectos de la radiaciónRESUMEN
In this study the hypothesis is tested that the motion pattern of small anatomic landmarks, recognizable at the left ventricular endocardial border in the contrast angiocardiogram, reflects the motion of the endocardial wall. To verify this, minute metal markers were inserted in the endocardium of eight pigs with a novel retrograde transvascular approach. Marker motion was subsequently recorded with roentgen cinematography and compared with the motion of the landmarks on the endocardial contours detected from the contrast ventriculogram with an automated contour detection system. Linear regression analysis of the directions of the systolic metal marker and endocardial landmark pathways yielded a correlation coefficient of 0.86 and a standard error of the estimate of 10.3 degrees. Landmark pathways were also measured in 23 normal human left ventriculograms. Normal left ventricular endocardial wall motion during systole, as observed in the 30 degrees right anterior oblique view, is characterized by a dominant inward transverse motion of the opposite anterior and inferoposterior walls and a descent of the base toward the apex. The apex itself is almost stationary. On the basis of these observations, a widely applicable model for the assessment of left ventricular wall motion is described in mathematical terms.
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Endocardio/anatomía & histología , Función Ventricular , Animales , Cineangiografía , Diástole , Endocardio/fisiología , Ventrículos Cardíacos/anatomía & histología , Ventrículos Cardíacos/diagnóstico por imagen , Humanos , Modelos Cardiovasculares , Movimiento , Estadística como Asunto , Porcinos , SístoleRESUMEN
OBJECTIVE: To compare percutaneous transluminal coronary angioplasty (PTCA) and stent implantation with respect to the long-term changes they induce in the newly formed endothelium in porcine coronary arteries by studying both morphological and functional parameters of the endothelium at 2 weeks and 3 months after intervention. BACKGROUND: Problems affecting PTCA or stent implantation have been overcome to a large extent by means of better techniques and the availability of new drugs. Late problems, however, still exist in that restenosis affects a large number of patients. With an increasing number of patients being treated with stents, the problem of in-stent restenosis is of even greater concern, as this seems difficult to treat. A functional endothelial lining is thought to be important in controlling the growth of the underlying vascular tissue. We hypothesized that the enhanced neointimal hyperplasia observed after stenting is associated with a more pronounced and prolonged endothelial dysfunction. METHODS: Arteries were analyzed using a dye-exclusion test and planimetry of permeable areas. Thereafter, the arteries were processed for light and scanning electron microscopy for assessment of morphology and proliferative response. RESULTS: Leakage of the endothelium for molecules such as Evans blue-albumin as well as prolonged endothelial proliferation is observed as late as 3 months after the intervention, and is more pronounced after stenting. Permeability is associated with distinct morphologic characteristics: endothelial retraction, the expression of surface folds, and the adhesion of leukocytes. CONCLUSIONS: Stenting especially decreases long-term vascular integrity with respect to permeability and endothelial proliferation, and is associated with distinct morphologic characteristics.
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Angioplastia Coronaria con Balón/efectos adversos , Vasos Coronarios/patología , Endotelio Vascular/patología , Stents/efectos adversos , Animales , Permeabilidad Capilar , División Celular , Vasos Coronarios/fisiopatología , Vasos Coronarios/ultraestructura , Endotelio Vascular/fisiopatología , Endotelio Vascular/ultraestructura , Porcinos , Túnica Íntima/fisiopatologíaRESUMEN
A 15 min reduction in blood flow in the left anterior descending coronary artery to 35% of baseline in open chest anaesthetised pigs produced a 25% decrease in cardiac output and a similar fall in maximum left ventricular dP/dt, whereas left ventricular filling pressure increased from 10(1) to 14(1) mmHg. The decrease in perfusion of the myocardial area nourished by the left anterior descending coronary artery was not evenly distributed since the endocardial to epicardial flow ratio decreased from 0.93(0.07) to 0.48(0.06). Regional myocardial wall thickening of the ischaemic segment decreased from 0.36(0.03) to 0.14(0.03), whereas the arterial-coronary venous differences in pH and PCO2 tripled. Subsequently, eight animals received 1.5 mg X kg-1 of the cardioselective beta adrenoceptor antagonist bevantolol, whereas seven other animals were treated with the solvent. In the following 15 min the cardiovascular performance of the solvent treated animals did not change appreciably, although there was a tendency to further deterioration. Bevantolol did not improve transmural myocardial blood flow to the ischaemic zone but caused a redistribution in favour of the endocardial layers since the endocardial to epicardial flow ratio almost returned to baseline. These changes in flow were accompanied by a narrowing of the arterial-coronary venous differences in pH and PCO2, but regional myocardial function did not improve. In another series of experiments, however, bevantolol (1.5 mg X kg-1) decreased the velocity of regional wall thickening of normal myocardium by 22(2)% as a result of its negative inotropic properties. Thus bevantolol appears to alleviate ischaemia, and the increase in diastolic perfusion time (17%) may be one of the major mechanisms.
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Antagonistas Adrenérgicos beta/uso terapéutico , Enfermedad Coronaria/tratamiento farmacológico , Propanolaminas/uso terapéutico , Animales , Modelos Animales de Enfermedad , Evaluación de Medicamentos , Corazón/efectos de los fármacos , Hemodinámica/efectos de los fármacos , PorcinosRESUMEN
OBJECTIVE: The present study determined the role of the autonomic nervous system (ANS) in the regulation of systemic and pulmonary circulation and of O2 delivery and utilization in swine at rest and during graded treadmill exercise. METHODS: Instrumented swine (n = 12) were subjected to treadmill exercise (1-5 km/h) under control conditions and in the presence of single and combined beta-adrenergic, alpha-adrenergic and muscarinic (M) receptor blockade. RESULTS: Exercise produced a four-fold increase in body O2 consumption, due to a doubling of both cardiac output and the arterio-mixed-venous O2 content difference. The latter resulted from an increase in O2 extraction, from 45 +/- 1% at rest to 74 +/- 1% at 5 km/h, as the O2 carrying capacity [haemoglobin concentration (Hb)] increased by only approximately 10%. The increase in cardiac output resulted from a doubling of the heart rate and a small (< 10%) increase in stroke volume. The mean aortic pressure (MAP) was unchanged, implying a 50% decrease in systemic vascular resistance (P < or = 0.05). In contrast, exercise had no significant effect on pulmonary vascular resistance. The sympathetic division of the ANS controlled O2 delivery via beta-adrenoceptors (heart rate and contractility) and Hb concentration via alpha-adrenoceptor-mediated splenic contraction. In addition, the sympathetic division modulated systemic vascular tone via alpha- and beta-adrenoceptors, but also exerted a vasodilator influence on the pulmonary circulation via beta-adrenoceptors. The parasympathetic division controlled O2 delivery in part directly (heart rate) and in part indirectly via inhibition of beta-adrenoceptor activity (heart rate and contractility), even during heavy exercise. In addition, the parasympathetic division exerted a direct vasodilator influence on the pulmonary, but not on the systemic, circulation. CONCLUSIONS: Thus, in swine, in a manner similar to that in humans, both the sympathetic and parasympathetic division of the ANS contribute to cardiovascular homeostasis during exercise up to levels of high intensity.
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Sistema Nervioso Autónomo/fisiología , Circulación Sanguínea/fisiología , Fenómenos Fisiológicos Cardiovasculares , Oxígeno/metabolismo , Esfuerzo Físico/fisiología , Circulación Pulmonar/fisiología , Porcinos/fisiología , Antagonistas Adrenérgicos alfa/farmacología , Antagonistas Adrenérgicos beta/farmacología , Análisis de Varianza , Animales , Atropina/farmacología , Dióxido de Carbono/sangre , Fenómenos Fisiológicos Cardiovasculares/efectos de los fármacos , Femenino , Concentración de Iones de Hidrógeno , Masculino , Antagonistas Muscarínicos/farmacología , Presión Parcial , Fentolamina/farmacología , Propranolol/farmacologíaRESUMEN
OBJECTIVE: The role of nitric oxide (NO) in the regulation of vasomotor tone and tissue O(2)-consumption is incompletely understood. We therefore determined the contribution of endogenous NO to regulation of systemic, pulmonary and coronary vasomotor tone and myocardial (MV(O(2))) and whole body (BV(O(2))) O(2)-consumption in exercising swine. METHODS AND RESULTS: Exercise (1-5 km/h) up to 85% of maximum heart rate in 11 swine produced a 4-fold increase in BV(O(2)), which was accommodated for by 2-fold increases in both cardiac output (CO) and body O(2)-extraction. The NO synthase inhibitor N(omega)-nitro-L-arginine (NLA, 20 mg/kg, i.v.) increased mean aortic pressure by 30 mmHg both at rest and during exercise, due to a decrease in systemic vascular conductance from 37+/-2 to 22+/-1 ml/min mmHg(-1) at rest and from 88+/-3 to 60+/-3 ml/min mmHg(-1) at 5 km/h (all P< or =0.05 versus control). NLA produced vasoconstriction at rest and at 5 km/h in virtually all regional beds but did not affect the exercise-induced redistribution of CO. NLA increased mean pulmonary artery pressure from 15+/-1 to 21+/-1 mmHg at rest and from 30+/-2 to 40+/-2 mmHg at 5 km/h, due to a decrease in pulmonary vascular conductance (all P< or =0.05). BV(O(2)) remained unchanged and consequently the decrease in CO resulted in a compensatory increase in O(2)-extraction. NLA in a dose of 40 mg/kg produced similar responses. NLA had no significant effect on myocardial O(2)-demand or MV(O(2)) either at rest or during exercise, but decreased coronary vascular conductance which resulted in a decrease in coronary venous PO(2) from 24.5+/-1.1 to 21.9+/-0.8 mmHg at rest and from 23.5+/-0.5 to 21.0+/-0.6 mmHg at 5 km/h (all P< or =0. 05). CONCLUSIONS: Endogenous NO dilates the systemic, pulmonary and coronary vascular bed, but does not modify MV(O(2)) or BV(O(2)) in swine at rest and during exercise.
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Ejercicio Físico/fisiología , Miocardio/metabolismo , Óxido Nítrico/fisiología , Consumo de Oxígeno/efectos de los fármacos , Resistencia Vascular/efectos de los fármacos , Animales , Circulación Coronaria/efectos de los fármacos , Perros , Inhibidores Enzimáticos/farmacología , Femenino , Humanos , Masculino , Músculo Esquelético/irrigación sanguínea , Óxido Nítrico Sintasa/antagonistas & inhibidores , Nitroarginina/farmacología , Circulación Pulmonar/efectos de los fármacos , Flujo Sanguíneo Regional/efectos de los fármacos , Especificidad de la Especie , Porcinos , Vasoconstrictores/farmacologíaRESUMEN
The effect of intravenously administered aprindine (AC1802) as a prophylactic agent against ventricular arrhythmias was studied in pigs. During the first 30 min of ischaemia 5 of the 22 untreated animals died because of ventricular fibrillation against 1 of the 23 animals pretreated with aprindine (P=0.09). Ventricular tachycardias were observed in 10 untreated animals and in none of the aprindine group (P=0.0002). The incidence of other arrhythmias was significantly less in the aprindine group compared with the untreated group (P less than 0.02).
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Aprindina/administración & dosificación , Arritmias Cardíacas/prevención & control , Indenos/administración & dosificación , Animales , Dióxido de Carbono/sangre , Enfermedad Coronaria/complicaciones , Porcinos , Fibrilación Ventricular/prevención & controlRESUMEN
OBJECTIVE: In order to determine whether the relatively high oxygen consumption of stunned myocardium is related to decreased mechanical efficiency, myocardial oxygen consumption (MVO2) and its major determinants were studied in 10 open chest anaesthetised pigs. METHODS: According to the time varying elastance concept, MVO2 is determined by contractility (Emax) and total mechanical work (PLA), which is the sum of the external work (EW) and potential energy (PE). Mechanical efficiency (EW/MVO2) equals the product of EW/PLA (= efficiency of energy transfer or EET) and PLA/MVO2. Emax is the slope of the end systolic pressure-segment length relationship, determined by gradually clamping the aorta. PLA is the area enclosed by the end systolic pressure-segment length relationship and the pressure-segment length trajectory. EW is the area of the pressure-segment length loop. Systemic haemodynamics, regional segment shortening, and MVO2 were determined at baseline, during stunning (two sequences of 10 min occlusion and 30 min of reperfusion), after a subsequent 50 beats.min-1 increase in heart rate by atrial pacing and additional infusion of 2 micrograms.kg-1.min-1 dobutamine. RESULTS: Stunning decreased segment shortening from 18.2(SEM 1.9)% to 10.2(1.5)%, MVO2 from 4.16(0.27) x 10(-2) to 2.84(0.25) x 10(-2) mumol.beat-1.g-1, and Emax from 47(9) to 23(3) mm Hg.mm-1 (all p < 0.05). PLA decreased by 13(4)%, as EW decreased by 42(6)%, and PE tended to increase. Although EET decreased from 0.58(0.04) to 0.40(0.03) (p < 0.05), there was no decrease in the mechanical efficiency, as an increase in PE caused an increase in PLA/MVO2 which compensated for the decrease in EET. Dobutamine infusion increased Emax and EW per beat to 120(23)% and 67(8)% of baseline, respectively, while MVO2 [4.12(0.53) mumol.beat-1.g-1] and EET [0.57(0.04)] returned to baseline. CONCLUSIONS: In stunned myocardium, mechanical efficiency is not decreased despite a decrease in EET. The increase in EET after dobutamine may explain the lack of the excessive increase in MVO2.
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Dobutamina/farmacología , Metabolismo Energético/efectos de los fármacos , Daño por Reperfusión Miocárdica/metabolismo , Miocardio/metabolismo , Animales , Modelos Animales de Enfermedad , Femenino , Corazón/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Masculino , Contracción Miocárdica/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Estimulación Química , Volumen Sistólico/fisiología , PorcinosRESUMEN
OBJECTIVE: The aim was to investigate whether ischaemic preconditioning can be obtained by a partial coronary artery occlusion without intermittent reperfusion. METHODS: In seven anaesthetised open chest pigs, the flow in the proximal left anterior descending coronary artery was reduced to 30% of baseline during 30 min before the vessel was occluded completely for 60 min (60 min total coronary occlusion, TCO). After 2 h of reperfusion, the area at risk (AR) and infarct size (IS) were determined using standard procedures. Infarct sizes were compared to those observed in control animals (n = 12), which were subjected to 60 min TCO and 2 h reperfusion, and to infarct sizes determined in animals preconditioned by 10 min TCO with either 15 min (n = 10) or 60 min (n = 5) of reperfusion before the 60 min TCO and 2 h reperfusion. In the last three groups of animals, area at risk was varied by occluding the coronary artery or its branches at different sites. RESULTS: In the control animals infarct size was linearly related (r = 0.99, p < 0.001) to the area at risk with a positive intercept on the AR axis: IS/LVmass (x100%) = 0.88 AR/LVmass (x100%)-3.6. At comparable areas at risk, the infarct size of the animals preconditioned with a 10 min TCO was less than for the control animals. For the animals preconditioned with 10 min TCO and 15 min reperfusion, the relationship between infarct size and area at risk was again linear (r = 0.88) and also had a positive intercept on the AR axis: IS/LVmass (x100%) = 0.68 AR/LVmass (x100%)-4.8. All animals with the flow reduction to 30% of baseline immediately preceding the 60 min TCO had infarct sizes smaller (p < 0.05) than predicted from the regression equation for the control animals, but the infarct size limitation could not be simply related to variables such as changes in regional systolic and postsystolic segment length shortening, ATP, or ADP during the partial occlusion period. CONCLUSIONS: Myocardium can be preconditioned with a flow reduction to 30% of baseline for 30 min without intermittent reperfusion (two stage Harris model). The positive intercept on the AR axis of the IS-AR relationship warrants caution of the use of IS/AR as an index for infarct size limitation.
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Vasos Coronarios , Infarto del Miocardio/prevención & control , Isquemia Miocárdica , Animales , Arteriopatías Oclusivas , Constricción , Modelos Animales de Enfermedad , Infarto del Miocardio/patología , Isquemia Miocárdica/patología , Miocardio/patología , PorcinosRESUMEN
A radioisotope procedure was used to determine long chain fatty acylcarnitine concentrations in fractions of porcine myocardium that had been subjected to different periods of ischaemia (0, 1, 2, and 3 h). In myocardial tissue from non-ligated hearts acylcarnitine concentrations were 0.32(0.03) and 1.53(0.04) nmol.mg-1 protein for homogenate and sarcolemma enriched membrane respectively, which indicates a preferential membrane localisation of long chain fatty acylcarnitine. Both the total and membrane acylcarnitine contents were increased about twofold after 2 h of ischaemia. The accumulation of long chain fatty acylcarnitine was not correlated temporally with changes in adenosine triphosphate dependent calcium uptake activity of homogenates (mainly a function of sarcoplasmic reticulum membranes), sodium gradient induced calcium uptake, and calcium permeability of sarcolemma preparations. Homogenate adenosine triphosphate dependent calcium uptake was decreased by 36% after 3 h of ischaemia. Sodium gradient induced calcium uptake was enhanced about twofold after 1 h of ischaemia, and calcium permeability of sarcolemmal vesicles was decreased by 20% after 3 h of ischaemia. After in vitro incubation of isolated sarcolemma membranes with (1-14C)-palmitoylcarnitine radiolabelled molecules that remained associated with the membrane even after repeated washing were incorporated. No changes were observed in the sodium gradient induced calcium uptake when less than 6 nmol (1(-14)C)- palmitoylcarnitine per mg sarcolemma protein were bound or incorporated into the lipid phase. This exceeded the maximal endogenous concentrations of 3.2(0.6) nmol long chain fatty acylcarnitine per mg sarcolemma protein observed during myocardial ischaemia. The results suggest that the intracellular increase in long chain acylcarnitine during almost zero myocardial flow is not critical to sarcolemmal sodium and calcium permeability and sarcoplasmic reticulum calcium pumping activity.
Asunto(s)
Acetilcarnitina/metabolismo , Calcio/metabolismo , Carnitina/análogos & derivados , Permeabilidad de la Membrana Celular , Enfermedad Coronaria/metabolismo , Miocardio/metabolismo , Animales , Miocardio/análisis , Palmitoilcarnitina/metabolismo , Sarcolema/metabolismo , PorcinosRESUMEN
OBJECTIVE: The aim was to evaluate the applicability of myocardial contrast echocardiography in the measurement of coronary flow reserve. METHODS: Eleven anaesthetised open chest pigs were studied, in which coronary atherosclerosis had been induced by abrasion of the left anterior descending coronary artery at one month, followed by an atherogenic diet for eight months. Coronary flow reserve was determined by electromagnetic flow measurement and contrast echocardiography before and after partial occlusion of the left anterior descending coronary artery, using papaverine as a coronary vasodilator. Coronary blood flow was reduced by tightening a clamp placed around the coronary artery. Systemic haemodynamics and myocardial wall thickness (epicardial ultrasound 5 MHz transducer) were recorded simultaneously. Echocardiograms were recorded on VHS tape and analysed by digitised videodensitometry off line for construction of the time v videointensity curve (time-intensity curves). From these curves washout time (T50), area under the curve, peak contrast intensity, and time to peak intensity were calculated. RESULTS: Following papaverine, coronary blood flow increased significantly from 47 (SD 23) ml.min-1 at baseline to 88(39) ml.min-1 (p less than 0.05). During the stenosis, flow decreased to 19(16) ml.min-1 (p less than 0.01), and increased to 38(29) ml.min-1 (p less than 0.05 v stenosis) after administration of papaverine. Correlations between coronary blood flow and indices calculated from the quantitative videodensitometric analysis were poor, varying between r = 0.03 for area at control flow to r = 0.62 for T50 during stenosis. The same was true for coronary flow reserve: r = 0.09 for peak to r = 0.75 (p less than 0.05) for time to peak without the stenosis. CONCLUSIONS: Current limitations in injection, imaging, and analysis techniques cause variability in data from time-intensity curves, which precludes accurate quantification of coronary flow (reserve) by myocardial contrast echocardiography.
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Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Circulación Coronaria/fisiología , Ecocardiografía/métodos , Animales , Presión Sanguínea/fisiología , Medios de Contraste , Modelos Animales de Enfermedad , Fenómenos Electromagnéticos , Papaverina/farmacología , Flujo Sanguíneo Regional/efectos de los fármacos , Flujo Sanguíneo Regional/fisiología , Porcinos , Factores de TiempoRESUMEN
OBJECTIVES: Previous studies have demonstrated a decreased flow reserve in the surviving hypertrophied left ventricle (LV) early after myocardial infarction. We hypothesized that exacerbation of hemodynamic abnormalities and neurohumoral activation during exercise could exhaust coronary flow reserve and thereby impair myocardial O(2) supply. Consequently, we studied hemodynamic, neurohumoral and regional myocardial perfusion and metabolic responses to exercise in pigs with LV hypertrophic remodeling 3 weeks after a myocardial infarction produced by permanent left circumflex coronary artery ligation. METHODS: Chronically instrumented pigs were exercised on a treadmill up to 85% of maximum heart rate. Pigs with a myocardial infarction (MI) had a lower cardiac output (21%), stroke volume (28%), LVdP/dt(max) (18%), systemic (22%) and pulmonary (20%) vascular conductance, and increased left atrial (225%) and pulmonary artery (75%) pressures, compared to normal pigs. In MI, the exercise-induced increases in cardiac pump function, and systemic and pulmonary vasodilation were blunted compared to normals. Consequently, perfusion of visceral organs became impaired during strenuous exercise, but cerebral and skeletal muscle blood flows were maintained. Exercise-induced increases in norepinephrine and endothelin levels were exacerbated and, while relative sympathetic drive was maintained, cardiac responsiveness to norepinephrine was blunted. Despite lower capillary densities in the hypertrophied non-infarcted LV and relative subendocardial hypoperfusion during strenuous exercise, which necessitated a slight increase in O(2) extraction, there was no metabolic evidence of overt myocardial ischemia during strenuous exercise as indicated by the arterio-coronary venous pH difference. CONCLUSIONS: LV dysfunction and neurohumoral activation were present in pigs with a 3-week-old infarction, particularly during exercise. However, although myocardial perfusion and O(2) supply were slightly impaired, myocardial ischemia did not occur even during exercise up to 85% of maximum heart rate, suggesting that perfusion abnormalities do not contribute to LV dysfunction early after infarction.
Asunto(s)
Circulación Coronaria , Infarto del Miocardio/fisiopatología , Condicionamiento Físico Animal/fisiología , Remodelación Ventricular , Animales , Encéfalo/irrigación sanguínea , Epinefrina/sangre , Femenino , Hemodinámica , Masculino , Músculo Esquelético/irrigación sanguínea , Infarto del Miocardio/metabolismo , Norepinefrina/sangre , Consumo de Oxígeno , Circulación Pulmonar , Porcinos , Factores de Tiempo , Resistencia VascularRESUMEN
STUDY OBJECTIVE: The aim as to compare the responses of intracoronary infusions of ATP, an endothelium dependent vasodilator, with adenosine following brief ischaemia (10 min) and reperfusion in a model of myocardial stunning. DESIGN: In group 1 (n = 6), coronary blood flow and endocardial (endo) and epicardial (epi) percent segment length shortening were measured in the distribution of the left anterior descending coronary artery before and during maximal intracoronary infusions of either adenosine or ATP (20 micrograms.kg-1.min-1). Measurements were obtained before and after myocardial stunning both at control heart rate and during atrial pacing (150 beats.min-1). In group 2 (n = 6), myocardial blood flows by microspheres and arterial-venous lactate and oxygen differences were determined following the same ischaemia-reperfusion protocol to characterise transmural changes in blood flow and metabolism in this model of stunning. EXPERIMENTAL MATERIAL: The experiments were done on 12 anaesthetised swine, weight 25-39 kg. MEASUREMENTS AND MAIN RESULTS: In group 1, baseline endo and epi segment length shortening were 16(SD 3)% and 14(6)% and following reperfusion were reduced to 10(4)% and 8(6)% respectively (p less than 0.05). Prior to stunning, minimal coronary resistances during adenosine and ATP were 0.81(0.40) and 0.76(0.25) mm Hg.min.ml-1 respectively and following reperfusion were 0.86(0.31) (NS) and 0.85(0.23) (NS) mm Hg.min.ml-1 respectively. Infusion of either vasodilator enhanced function by 30% following reperfusion whereas no such effect was observed prior to ischaemia. In group 2, no maldistribution of blood flow was observed following the same ischaemia-reperfusion protocol to account for this vasodilator enhancement in function. Percent lactate extraction values were 29(11)% and 25(14)% at preischaemic control and paced heart rates respectively, and following reperfusion were lowered to 0(12)% without pacing (p less than 0.05) and -1(34)% during pacing (p less than 0.05). CONCLUSIONS: Brief ischaemia and reperfusion in swine induces myocardial stunning without altering the vasodilator responses of either ATP, an endothelium dependent vasodilator, or adenosine. Recruitment in postischaemic segment length shortening was observed during infusions of both vasodilators at a time when maldistribution of flow was not observed. Possible mechanisms include either enhanced washout of lactate from the reperfused myocardium or greater utilisation of substrates during higher blood flows.
Asunto(s)
Adenosina Trifosfato/farmacología , Enfermedad Coronaria/fisiopatología , Vasos Coronarios/efectos de los fármacos , Endotelio Vascular/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Adenosina/farmacología , Animales , Vasos Coronarios/fisiopatología , Modelos Animales de Enfermedad , Endotelio Vascular/fisiopatología , Femenino , Masculino , Reperfusión Miocárdica/métodos , Marcapaso Artificial , Flujo Sanguíneo Regional/efectos de los fármacos , Porcinos , Resistencia Vascular/efectos de los fármacosRESUMEN
OBJECTIVE: Previous observations have shown that myocardium activates many adaptive processes after brief ischaemia. The aim of this study was to determine whether insulin-like growth factors (IGF) as well as their receptors and binding proteins (IGFBP), which control the activity of the IGF, may play an important role during these processes. METHODS: Ischaemia was induced in anaesthetised open chest pigs by two 10 min occlusions of the left anterior descending coronary artery, separated by 30 min of reperfusion, and followed by reperfusion up to 210 min. Tissue from the ischaemic area and from a non-ischaemic control region of the same heart was examined by means of northern blot, slot blot, and in situ hybridisation. RESULTS: IGF-I, IGF-II, the type I receptor, the insulin receptor, and IGFBP-2-6 are constitutively expressed in porcine myocardium. In situ hybridisation showed that IGF-I and IGF-II are mainly transcribed by myocytes. Ischaemia/reperfusion led to an early and significant increase in IGF-II mRNA compared to non-sham controls but not in comparison with sham operated animals, which already showed a (not significantly) enhanced IGF-II expression. In each case the IGF-II mRNA levels are equal in the control and the experimental region of the same heart. Whereas IGF-II expression was already increased by experimental stress, IGFBP-5 mRNA was enhanced only by ischaemia/reperfusion. The expression of IGF-I, the receptors, and IGFBP-2, 3, 4, and 6 remained unchanged during the experimental protocol. IGFBP-1 was neither expressed nor induced in our model. CONCLUSIONS: IGF-II acts like a stress-response gene activated by the experimental conditions (surgery, anaesthesia) and remains induced during following episodes of ischaemia/reperfusion. A possible interaction of IGFBP-5 with other components of the IGF system may contribute to the preconditioning response.
Asunto(s)
Regulación de la Expresión Génica , Factor II del Crecimiento Similar a la Insulina/genética , Isquemia Miocárdica/metabolismo , Miocardio/metabolismo , Animales , Northern Blotting , Proteínas Portadoras/genética , Inhibidores de Crecimiento/genética , Hibridación in Situ , Proteína 5 de Unión a Factor de Crecimiento Similar a la Insulina , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina , Isquemia , Reperfusión Miocárdica , ARN Mensajero/análisis , PorcinosRESUMEN
OBJECTIVE: Recently, we reported that a partial coronary artery occlusion immediately preceding a sustained coronary artery occlusion limited infarct size. We now investigated whether the protection by partial coronary artery occlusions (i) depends on the severity and(or) duration of the flow reduction and (ii) varies in the different myocardial layers. METHODS: In 71 open-chest pigs (eight groups) left ventricular area at risk (AR) and infarct area (IA) were determined for the endocardial (IAendo and ARendo) and epicardial halves (IAepi and ARepi). RESULTS: In control animals (60 min total coronary artery occlusion (TCO) followed by 120 min reperfusion (Rep)) there were highly linear relations between IA and AR in the endocardium (r = 0.98, P < 0.01) and epicardium (r = 0.97, P < 0.01), which could be described by IAendo = 1.01 ARendo - 4.5 and by IAepi = 0.88ARepi - 3.6, respectively. In animals that underwent a 10 min TCO + 15 min Rep prior to the 60 min TCO + 120 min Rep, IA in both myocardial layers were again highly linearly related with AR, with less steep slopes for both the endocardium (0.63) and epicardium (0.57) (both P < 0.01). Two groups of pigs were subjected to either a 30 or 90 min 70% reduction in coronary blood flow (FR) immediately preceding the 60 min TCO + 120 min Rep, without intervening reperfusion. A 30 min 70% FR decreased IA to the same degree in the endo- and epicardial half. A 90 min 70% FR resulted in protection in the epicardium (P < 0.01) but not in the endocardium, most likely because 90 min 70% FR without 60 min TCO already caused infarction which was more severe in the endo- than in the epicardium (P < 0.01). Endocardial and epicardial IA after either a 30 or 90 min 30% FR prior to the 60 min TCO was not different from that in the control group, indicating that this mild flow reduction failed to limit irreversible damage. CONCLUSIONS: Thirty or ninety min of severe (70%) but not mild (30%) coronary flow reductions protected against myocardial infarction. The protection by a 70% FR was influenced by the duration of FR as a 30 min 70% FR similarly decreased IA in the endocardial and epicardial halves, while 90 min 70% FR preferentially limited IA in the epicardial half. These findings suggest that perfusion abnormalities immediately preceding an infarction could be an important source of infarct size variability in patients.
Asunto(s)
Infarto del Miocardio/prevención & control , Isquemia Miocárdica/patología , Miocardio/patología , Animales , Circulación Coronaria , Endocardio/patología , Microscopía Fluorescente , Contracción Miocárdica , Infarto del Miocardio/patología , Pericardio/patología , Estadísticas no Paramétricas , Porcinos , Factores de TiempoRESUMEN
OBJECTIVE: Oxygen consumption (MVO2) of stunned myocardium is relatively high compared to, and poorly correlated with, systolic contractile function. The aim of this study was to investigate whether an increased afterload dependency, induced by the decreased contractility of the stunned myocardium, contributes to the large variability in the mechanical efficiency data. METHODS: In 13 anaesthetised open thorax pigs undergoing two cycles of 10 min occlusion of left anterior descending coronary artery and 30 min reperfusion, segment shortening, the slope of end systolic pressure segment length relationship (Ees), external work (EW, derived from the area inside the left ventricular pressure segment length loop), the efficiency of energy conversion (EET, = EW/PLA x 100%, where PLA = total pressure-segment length area), mechanical efficiency (EW/MVO2), and their dependency on left ventricular end systolic pressure (Pes) were determined before and after induction of stunning, and during subsequent inotropic stimulation with dobutamine (1 and 3 micrograms.kg-1.min-1 over 15 min). RESULTS: The stunning protocol not only caused significant decreases in segment shortening, external work, energy conversion efficiency, and EW/MVO2 but also increased the afterload dependency of these variables. Before stunning an increase in Pes from 100 to 160 mm Hg decreased segment shortening from 18(SEM 1)% to 14(2)% (P > 0.05) and increased external work from 206(18) to 254(32) mm Hg.mm (P < 0.05). After induction of stunning the same increase in Pes caused a decrease in segment shortening from 9.5(1.8)% to -4.6(2.1)% (P < 0.05) and in external work from 149(21) to -11(10) mm Hg.mm (P < 0.05). The afterload dependency of the PLA was not altered by stunning, but the afterload dependency of energy conversion efficiency increased, since efficiency decreased from 67(3)% to 59(5)% as Pes was increased from 100 to 160 mm Hg before stunning, but from 57(5) to -7(5)% after induction of stunning (P < 0.05). Furthermore, the same increase in Pes resulted in an 8% decrease of EW/MVO2 before stunning and 107% after induction of stunning. Infusion of dobutamine not only restored segment shortening, external work, energy conversion efficiency, and EW/MVO2 of the stunned myocardium, but also attenuated their afterload dependency to pre-stunning levels. CONCLUSIONS: Myocardial stunning increases the afterload dependency of segment shortening, external work, energy conversion efficiency, and mechanical efficiency, which can be attenuated by inotropic stimulation with dobutamine. However, the decrease in left ventricular end systolic pressure, which accompanies the induction of stunning, counteracts the decrease in these variables. These two mechanisms can explain most of the reported scatter in mechanical efficiency.
Asunto(s)
Corazón/fisiopatología , Contracción Miocárdica/fisiología , Aturdimiento Miocárdico/fisiopatología , Animales , Dobutamina/farmacología , Metabolismo Energético/fisiología , Corazón/efectos de los fármacos , Porcinos , SístoleRESUMEN
OBJECTIVES: Characterization of an in vitro model of endothelin-1 induced hypertrophy of cultured neonatal rat ventricular myocytes and subsequent analysis of transcription regulation of the rat promoter of the sarcoplasmic reticulum Ca2+ ATPase gene. METHODS: Neonatal rat ventricular myocytes were cultured in serum free medium and hypertrophy was induced by addition of endothelin-1 to 10(-8) M up to 48 h. Hypertrophy was characterized biochemically, and gene expression regulation was evaluated by Northern blotting. A sarcoplasmic reticulum Ca2+ ATPase promoter fragment, isolated from a rat library was cloned in a reporter vector. Promoter activity during hypertrophy was assessed after transfection of the reporter plasmid to cultured cardiomyocytes. RESULTS: Stimulation with endothelin-1 resulted in increased cell size, as indicated by protein/DNA ratio as well as by augmented protein synthesis. When compared to angiotensin II or alpha 1-adrenergic agonist, endothelin-1 was the strongest inducer of hypertrophy (protein/DNA ratio) after 48 h of stimulation. Endothelin-1 induced hypertrophy was accompanied by a twofold increase in total RNA content per cell as well as to increased glyceraldehydephosphate dehydrogenase mRNA levels. The level of atrial natriuretic factor mRNA was increased more than twofold, relative to glyceraldehydephosphate dehydrogenase, while the expression of the sarcoplasmic reticulum Ca2+ pump and phospholamban genes was decreased (by 26 and 49%, respectively) after induction of hypertrophy by stimulation with endothelin-1. In the same model, a 1.9 kb sarcoplasmic reticulum Ca2+ pump gene promoter fragment (including 0.4 kb of the 5' UTR of the mRNA) directed down-regulation of the expression of the reporter gene to the same magnitude as endogenous Ca2+ pump mRNA relative to glyceraldehydephosphate dehydrogenase mRNA. However, absolute mRNA level per cell did not change for either the reporter gene or the endogenous Ca2+ pump. CONCLUSIONS: Endothelin-1 can induce phenotypic changes in cultured rat ventricular myocytes that are reminiscent of hypertrophy in vivo. In this model, a 1.9 kb sarcoplasmic reticulum Ca2+ pump promoter fragment directed gene expression of a reporter gene identical to the endogenous regulation of the Ca2+ pump. Furthermore, expression of the Ca2+ pump during hypertrophy was only downregulated when compared to (increased levels of) glyceraldehydephosphate dehydrogenase mRNA, but absolute Ca2+ ATPase mRNA amounts remained unchanged. This suggests that the Ca2+ pump promoter is not responding to the increase in transcriptional activity that accompanies hypertrophy.