RESUMEN
Sodium-hydrogen exchange inhibitors, such as cariporide, are potent cardioprotective agents, however, safety concerns have been raised about intravenously (i.v.) administered cariporide in humans. The aim of this study was to develop a preservation strategy that maintained cariporide's cardioprotective efficacy during heart transplantation while minimizing recipient exposure. We utilized a porcine model of orthotopic heart transplantation that incorporated donor brain death and 14 h static heart storage. Five groups were studied: control (CON), hearts stored in Celsior; CAR1, hearts stored in Celsior with donors and recipients receiving cariporide (2 mg/kg i.v.) prior to explantation and reperfusion, respectively; CAR2, hearts stored in Celsior supplemented with cariporide (10 mumol/L); GTN, hearts stored in Celsior supplemented with glyceryl trinitrate (GTN) (100 mg/L); and COMB, hearts stored in Celsior supplemented with cariporide (10 mumol/L) plus GTN (100 mg/L). A total of 5/5 CAR1 and 5/6 COMB recipients were weaned from cardiopulmonary bypass compared with 1/5 CON, 1/5 CAR2 and 0/5 GTN animals (p = 0.001). Hearts from the CAR1 and COMB groups demonstrated similar cardiac function and troponin release after transplantation. Supplementation of Celsior with cariporide plus GTN provided superior donor heart preservation to supplementation with either agent alone and equivalent preservation to that observed with systemic administration of cariporide to the donor and recipient.
Asunto(s)
Guanidinas/administración & dosificación , Trasplante de Corazón/métodos , Nitroglicerina/administración & dosificación , Preservación de Órganos/métodos , Sulfonas/administración & dosificación , Animales , Antiarrítmicos/administración & dosificación , Peso Corporal , Disacáridos/administración & dosificación , Electrólitos/administración & dosificación , Glutamatos/administración & dosificación , Glutatión/administración & dosificación , Histidina/administración & dosificación , Isquemia , Manitol/administración & dosificación , Preservación de Órganos/instrumentación , Soluciones Preservantes de Órganos/administración & dosificación , Daño por Reperfusión/prevención & control , Porcinos , Factores de Tiempo , Vasodilatadores/administración & dosificaciónRESUMEN
We compared the effects of hormone resuscitation (HR) with a norepinephrine-based protocol on cardiac function, hemodynamics and need for vasopressor support after brain death in a porcine model. Following brain death induction, animals were treated with norepinephrine and fluids for 3 h. In the following 3 h, they continued on norepinephrine and fluids (control) or received additional HR (triiodothyronine, methylprednisolone, vasopressin, insulin). Data were collected pre-brain death, 3 and 6 h post-brain death. At 6 h, median norepinephrine use was higher in controls (0.563 vs. 0 microg/kg/min; p < 0.005), with 6/8 HR animals weaned off norepinephrine compared with 0/9 controls. Mean arterial pressure was higher in HR animals at 6 h (74 +/- 17 vs. 54 +/- 14 mmHg; p < 0.05). Cardiac contractility was also significantly higher in HR animals at 6 h (stroke work index 1.777 vs. 1.494). After collection of 6 h data, all animals were placed on the same low dose of norepinephrine. At 6.25 h, HR animals had higher stroke work (3540 +/- 1083 vs. 1536 +/- 702 mL.mmHg; p < 0.005), stroke volume (37.2 +/- 8.2 vs. 21.5 +/- 9.8 mL; p < 0.01) and cardiac output (5.8 +/- 1.4 vs. 3.2 +/- 1.2 L/min; p < 0.005). HR in a porcine model of brain death reduces norepinephrine requirements, and improves hemodynamics and cardiac function. These results support the use of HR in the management of the brain-dead donor.
Asunto(s)
Muerte Encefálica , Corazón/fisiología , Hormonas/farmacología , Resucitación/métodos , Donantes de Tejidos , Animales , Corazón/efectos de los fármacos , Insulina/farmacología , Metilprednisolona/farmacología , Modelos Animales , Norepinefrina/farmacología , Porcinos , Triyodotironina/farmacología , Vasopresinas/farmacologíaRESUMEN
We previously reported that transgenic (TG) mice with cardiac-restricted alpha(1A)-adrenergic receptor (alpha(1A)-AR)-overexpression showed enhanced contractility, but no hypertrophy. Since chronic inotropic enhancement may be deleterious, we investigated if long-term, cardiac function and longevity are compromised. alpha(1A)-TG mice, but not their non-TG littermates (NTLs), showed progressive loss of left ventricular (LV) hypercontractility (dP/dt(max): 14,567+/-603 to 11,610+/-915 mmHg/s, P<0.05, A1A1 line: 170-fold overexpression; and 13,625+/-826 to 8322+/-682 mmHg/s, respectively, P<0.05, A1A4 line: 112-fold overexpression, at 2 and 6 months, respectively). Both TG lines developed LV fibrosis, but not LV dilatation or hypertrophy, despite activation of hypertrophic signaling pathways. Microarray and real time RT-PCR analyses revealed activation of matricellular protein genes, including those for thrombospondin 1, connective tissue growth factor and tenascin C, but not transforming growth factor beta1. Life-span was markedly shortened (mean age at death: 155 days, A1A1 line; 224 days, A1A4 line compared with NTLs: >300 days). Telemetric electrocardiography revealed that death in the alpha(1A)-AR TG mice was due to cardiac standstill preceded by a progressive diminution in QRS amplitude, but not by arrhythmias. The QRS changes and sudden death could be mimicked by alpha(1)-AR activation, and reversed preterminally by alpha(1)-AR blockade, suggesting a relationship to stress- or activity-associated catecholamine release. Thus, long-term augmentation of cardiac alpha(1A)-adrenergic drive leads to premature death and progressive LV fibrosis with reactivation of matricellular protein genes. To our knowledge this is the first evidence in vivo for a role of the alpha(1A)-AR in ventricular fibrosis and in pathological cardiac remodeling.
Asunto(s)
Fibrosis Endomiocárdica/genética , Regulación de la Expresión Génica/genética , Contracción Miocárdica/genética , Receptores Adrenérgicos alfa 1/genética , Remodelación Ventricular/genética , Animales , Fibrosis Endomiocárdica/patología , Expresión Génica/genética , Perfilación de la Expresión Génica/métodos , Ratones , Miocardio/patología , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Receptores Adrenérgicos alfa 1/biosíntesisRESUMEN
Activation of the alpha(1A)-adrenergic receptor (alpha(1A)-AR)/Gq pathway has been implicated as a critical trigger for the development of cardiac hypertrophy. However, direct evidence from in vivo studies is still lacking. To address this issue, transgenic mice with cardiac-targeted overexpression of the alpha(1A)-AR (4- to 170-fold) were generated, using the rodent alpha-myosin heavy chain promoter. Heterozygous animals displayed marked enhancement of cardiac contractility, evident from increases in dP/dt(max) (80%, P<0.0001), dP/dt(max)/LVP(inst) (76%, P<0.001), dP/dt(max):dP/dt(min) (104%, P<0.0001), and fractional shortening (33%, P<0.05). Moreover, changes in the dP/dt(max)-end-diastolic volume relationship provided load-independent evidence of a primary increase in contractility. Blood pressure and heart rate were largely unchanged, and there was a small increase in (-)norepinephrine-stimulated, but not basal, phospholipase C activity. Increased contractility was directly related to the level of receptor overexpression and could be completely reversed by acute alpha(1A)- but not beta-AR blockade. Despite the robust changes in contractility, transgenic animals displayed no morphological, histological, or echocardiographic evidence of left ventricular hypertrophy. In addition, apart from an increase in atrial natriuretic factor mRNA, expression of other hypertrophy-associated genes was unchanged. To our knowledge, these data provide the first in vivo evidence for an inotropic action of the alpha(1A)-AR.
Asunto(s)
Cardiomegalia , Expresión Génica/fisiología , Marcación de Gen , Contracción Miocárdica/fisiología , Receptores Adrenérgicos alfa 1/biosíntesis , Adenilil Ciclasas/metabolismo , Antagonistas de Receptores Adrenérgicos alfa 1 , Agonistas alfa-Adrenérgicos/farmacología , Antagonistas Adrenérgicos alfa/farmacología , Animales , Factor Natriurético Atrial/genética , Factor Natriurético Atrial/metabolismo , Presión Sanguínea/genética , Ecocardiografía , Electrocardiografía/efectos de los fármacos , Frecuencia Cardíaca/genética , Heterocigoto , Fosfatos de Inositol/metabolismo , Ratones , Ratones Transgénicos , Contracción Miocárdica/efectos de los fármacos , Cadenas Pesadas de Miosina/genética , Tamaño de los Órganos/genética , Especificidad de Órganos/genética , Regiones Promotoras Genéticas , ARN Mensajero/metabolismo , Receptores Adrenérgicos alfa 1/genética , Transgenes/fisiología , Fosfolipasas de Tipo C/metabolismoRESUMEN
Heterozygous mutations in the cardiac homeobox gene, NKX2-5, underlie familial cases of atrial septal defect (ASD) with severe atrioventricular conduction block. In this study, mice heterozygous for Nkx2-5-null alleles were assessed for analogous defects. Although ASD occurred only rarely, atrial septal dysmorphogenesis was evident as increased frequencies of patent foramen ovale and septal aneurysm, and decreased length of the septum primum flap valve. These parameters were compounded by genetic background effects, and in the 129/Sv strain, septal dysmorphogenesis bordered on ASD in 17% of Nkx2-5 heterozygotes. In a proportion of neonatal heterozygotes, as well as in adults with ASD, we found that the size of the foramen ovale was significantly enlarged and altered in shape, potentially exposing the normally thin septum primum to excessive hemodynamic forces. Therefore, defective morphogenesis of the septum secundum may be one contributing factor in the generation of patent foramen ovale, septal aneurysm, and certain ASDs. Mild prolongation of P-R interval in females and an increased frequency of stenotic bicuspid aortic valves were also features of the Nkx2-5 heterozygous phenotype. Our data demonstrate that the complex effects of Nkx2-5 haploinsufficiency in mice are weaker but convergent with those in humans. As in the mouse, the phenotype of human NKX2-5 mutations may be modulated by interacting alleles.
Asunto(s)
Defectos de los Tabiques Cardíacos/genética , Válvulas Cardíacas/anomalías , Heterocigoto , Proteínas de Homeodominio/genética , Mutación/genética , Factores de Transcripción , Proteínas de Xenopus , Alelos , Animales , Animales Recién Nacidos , Velocidad del Flujo Sanguíneo , Ecocardiografía , Electrocardiografía , Genes Homeobox , Defectos de los Tabiques Cardíacos/diagnóstico por imagen , Defectos de los Tabiques Cardíacos/patología , Defectos del Tabique Interatrial/diagnóstico por imagen , Defectos del Tabique Interatrial/genética , Defectos del Tabique Interatrial/patología , Válvulas Cardíacas/diagnóstico por imagen , Válvulas Cardíacas/patología , Proteína Homeótica Nkx-2.5 , Ratones , Ratones Endogámicos , Ratones Transgénicos , Válvula Mitral/anomalías , Válvula Mitral/diagnóstico por imagen , Válvula Mitral/patología , Estenosis de la Válvula Mitral/diagnóstico por imagen , Estenosis de la Válvula Mitral/genética , Estenosis de la Válvula Mitral/patologíaRESUMEN
BACKGROUND: We aimed to develop a large animal model of orthotopic cardiac transplantation, incorporating donor brain death, to assess new methods of preservation of the donor heart. METHODS: Brain death was achieved in the donor pig by inflation of a 20 cc subdural balloon 1 h prior to harvest. The donor heart was stored for 6 h with conventional hypothermic ischaemic preservation. It was then transplanted orthotopically into the recipient pig using the Lower and Shumway technique. One hour after reperfusion, the transplanted heart was weaned from cardiopulmonary bypass with dobutamine support. Dobutamine support was continued for up to 4 h, if required. After 6 h of physiological and biochemical evaluation, the recipient was euthanased and the heart excised for histological assessment. RESULTS: All pigs experienced the classical haemodynamic changes associated with brain death. This resulted in the release of Troponin I, consistent with myocardial injury. The donor operation was successfully completed in 11 out of 13 pigs. Six out of 11 transplanted hearts were successfully weaned from cardiopulmonary bypass, but required ongoing dobutamine support. CONCLUSIONS: This porcine model of orthotopic cardiac transplantation is a relevant and practical large animal model for the assessment of new methods of preservation of the donor heart.
RESUMEN
BACKGROUND: Increased left ventricular (LV) contractile force or oxygen consumption has been documented with increased coronary arterial pressure (CAP) and flow (Gregg phenomenon). We investigated whether the increase in contractile force with increased LV afterload might be mediated by the concomitant increase in CAP when coronary autoregulation is intact. METHODS AND RESULTS: The LV of 6 autonomically blocked open-chest dogs was perfused through the left main coronary artery by a cannula with a side gate to the aortic root. With the gate open, CAP increased from 77+/-20 to 93+/-20 mm Hg (P<0.05) with aortic constriction (AC). With the gate closed, CAP was maintained at a constant level of 100 mm Hg. A small reduction in the slope of the preload recruitable stroke work (PRSW) relationship was observed with AC, but this response was not altered by the coronary perfusion gate position. The end-systolic pressure-volume (ESPV) relationship shifted upward significantly with AC (P<0.001), but this shift was not greater with open-gate perfusion than with closed-gate perfusion. Furthermore, with coronary autoregulation intact, wide changes in CAP (between 60 and 180 mm Hg, n=5) did not alter either the PRSW or ESPV relationship. In contrast, when autoregulation was abolished with intracoronary adenosine (n=6), both indexes of contractility increased progressively with increased CAP. CONCLUSIONS: The concomitant increase in CAP with increased afterload in the intact canine LV does not contribute to the afterload-induced increase in contractile force. Coronary perfusion pressure per se does not influence LV contractile function. Coronary perfusion pressure influences contractility only when coronary flow changes.
Asunto(s)
Presión Sanguínea , Circulación Coronaria , Contracción Miocárdica , Función Ventricular Izquierda , Animales , Perros , HomeostasisRESUMEN
1. Regional differences in odorant-induced responsiveness of the rat olfactory epithelium were measured via electrophysiological recordings [negative component of electro-olfactogram (Veog(-)) made from the surface of the olfactory epithelium on the nasal septum]. The nasal septum provided a flat surface from which multiple recordings could be made. 2. Veog(-)s were recorded from a standardized grid of 16 sites. This grid of recording sites extended over most of the surface of the olfactory epithelium on the nasal septum. 3. Twenty-one animals were tested for their responses to seven odorants. The animals were divided into three groups, each of which was tested with two different odorants plus amyl acetate, which provided a comparison between the groups. 4. For each odorant in each animal, topographic maps of relative responsiveness were derived to test whether odorants elicited different patterns of responses in the same individual. Topographic maps of responsiveness were derived also for the animal groups to test for the generality of the form of the maps for different odorants. Response latencies were also measured for each odorant at each recording site. 5. All individuals showed different topographic patterns of responses to the three test odorants. For most odorants, the location of the most responsive site was similar in all animals. In different animals the topographic maps for the same odorant were remarkably similar. Topographic maps for the odorants were all different from one another. 6. These results are consistent with the hypothesis that odorant quality is encoded in the differential spatial distribution of receptor cells whose differences in responsiveness appear to be distributed as a continuum across the epithelium. The results establish for a mammalian species what was previously reported in amphibians. These differences are presumed to be due to differential expression of odorant receptor proteins. 7. The mean response latency was 32 ms. This period was similar for all odorants, all animals, and all recording sites and was independent of Veog(-) amplitude. It is concluded that diffusion through the mucus contributed approximately 6 ms to the latency of onset of the responses to these odorants.