RESUMEN
Renal medullary endothelin B receptors (ET(B)) mediate sodium excretion and blood pressure (BP) control. Several animal models of hypertension have impaired renal medullary ET(B) function. We found that 4-week high-caloric diet elevated systolic BP in Dahl salt-sensitive (Dahl S) rats (126+/-2 vs. 143+/-3 mm Hg, p<0.05). We hypothesized that renal medullary ET(B) function is dysfunctional in DS rats fed a high-caloric diet. We compared the diuretic and natriuretic response to intramedullary infusion of ET(B) agonist sarafotoxin 6c (S6c) in DS rats fed either a normal or high-caloric diet for 4 weeks. Urine was collected during intramedullary infusion of saline for baseline collection followed by intramedullary infusion of either saline or S6c. We first examined the ET(B) function in DS rats fed a normal diet. S6c increased urine flow (2.7+/-0.3 microl/min during baseline vs. 5.1+/-0.6 microl/min after S6c; p<0.05; n=5) and sodium excretion (0.28+/-0.05 vs. 0.81+/-0.17 micromol/min; p<0.05), suggesting that DS rats have renal medullary ET(B) function. However, DS rats fed a high-caloric diet displayed a significant increase in urine flow (2.7+/-0.4 vs. 4.2+/-0.4 microl/min, baseline vs. S6c infusion, respectively; p<0.05, n=6), but no significant change in sodium excretion in response to S6c (0.32+/-0.06 vs. 0.45+/-0.10 micromol/min). These data demonstrate that renal medullary ET(B) function is impaired in DS rats fed a high-caloric diet, which may be contributed to the elevation of blood pressure during high-caloric feeding in this model.
Asunto(s)
Dieta Alta en Grasa/efectos adversos , Ingestión de Alimentos/fisiología , Hipertensión/metabolismo , Médula Renal/metabolismo , Receptor de Endotelina B/agonistas , Receptor de Endotelina B/metabolismo , Animales , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Ingestión de Alimentos/efectos de los fármacos , Hipertensión/etiología , Médula Renal/efectos de los fármacos , Masculino , Péptidos/farmacología , Ratas , Ratas Endogámicas Dahl , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Venenos de VíborasRESUMEN
AIM: This study was designed to determine whether ET-1 derived from endothelial cells contributes to oxidative stress in the glomerulus of mice subjected to a high-salt diet and/or hypoxia. METHODS: C57BL6/J control mice or vascular endothelial cell ET-1 knockout (VEET KO) mice were subjected to 3-h exposure to hypoxia (8% O2) and/or 2 weeks of high-salt diet (4% NaCl) prior to metabolic cage assessment of renal function and isolation of glomeruli for the determination of reactive oxygen species (ROS). RESULTS: In control mice, hypoxia significantly increased urinary protein excretion during the initial 24 h, but only in animals on a high-salt diet. Hypoxia increased glomerular ET-1 mRNA expression in control, but not in vascular endothelial cell ET-1 knockout (VEET KO) mice. Under normoxic conditions, mice on a high-salt diet had approx. 150% higher glomerular ET-1 mRNA expression compared with a normal-salt diet (P < 0.05). High-salt diet administration significantly increased glomerular ROS production in flox control, but not in glomeruli isolated from VEET KO mice. In C57BL6/J mice, the ETA receptor-selective antagonist, ABT-627, significantly attenuated the increase in glomerular ROS production produced by high-salt diet. In addition, chronic infusion of C57BL6/J mice with a subpressor dose of ET-1 (osmotic pumps) significantly increased the levels of glomerular ROS that were prevented by ETA antagonist treatment. CONCLUSION: These data suggest that both hypoxia and a high-salt diet increase glomerular ROS production via endothelial-derived ET-1-ETA receptor activation and provide a potential mechanism for ET-1-induced nephropathy.
Asunto(s)
Endotelina-1/administración & dosificación , Hipoxia/metabolismo , Enfermedades Renales/metabolismo , Glomérulos Renales/metabolismo , Estrés Oxidativo , Especies Reactivas de Oxígeno/metabolismo , Cloruro de Sodio Dietético/efectos adversos , Animales , Modelos Animales de Enfermedad , Antagonistas de los Receptores de la Endotelina A/farmacología , Endotelina-1/deficiencia , Endotelina-1/genética , Hipoxia/complicaciones , Enfermedades Renales/etiología , Enfermedades Renales/genética , Enfermedades Renales/fisiopatología , Glomérulos Renales/efectos de los fármacos , Glomérulos Renales/fisiopatología , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Estrés Oxidativo/efectos de los fármacos , Proteinuria/metabolismo , Proteinuria/fisiopatología , Receptor de Endotelina A/efectos de los fármacos , Receptor de Endotelina A/metabolismo , Cloruro de Sodio Dietético/metabolismo , Factores de TiempoRESUMEN
Early origins of adult disease may be defined as adversity or challenges during early life that alter physiological responses and prime the organism to chronic disease in adult life. Adverse childhood experiences or early life stress (ELS) may be considered a silent independent risk factor capable of predicting future cardiovascular disease risk. Maternal separation (MatSep) provides a suitable model to elucidate the underlying molecular mechanisms by which ELS increases the risk to develop cardiovascular disease in adulthood. The aim of this review is to describe the links between behavioural stress early in life and chronic cardiovascular disease risk in adulthood. We will discuss the following: (i) adult cardiovascular outcomes in humans subjected to ELS, (ii) MatSep as an animal model of ELS as well as the limitations and advantages of this model in rodents and (iii) possible ELS-induced mechanisms that predispose individuals to greater cardiovascular risk. Overall, exposure to a behavioural stressor early in life sensitizes the response to a second stressor later in life, thus unmasking an exaggerated cardiovascular dysfunction that may influence quality of life and life expectancy in adulthood.
Asunto(s)
Enfermedades Cardiovasculares/psicología , Estrés Psicológico/complicaciones , Animales , Humanos , Privación Materna , Factores de RiesgoRESUMEN
AIM: During type 1 diabetes (T1D), the medullary thick ascending limb (mTAL) displays an NADPH oxidase-dependent increase in sodium transport, in concert with increased NO production by NO synthase 1 (NOS1) and NOS2. We hypothesized that NOS1- and/or NOS2-derived NO blunts T1D-induced activation of sodium transport in the mTAL. METHODS: T1D was induced by streptozotocin injection (STZ rats); sham rats received vehicle. Three-to-four weeks later, mTAL were isolated from both groups for assay of nitrite and superoxide production, and O2 consumption in the absence or presence of various inhibitors. RESULTS: Apocynin (NADPH oxidase inhibitor) normalized superoxide production and ouabain-sensitive O2 consumption and furosemide-sensitive O2 consumption by mTALs from STZ rats, without altering O2 consumption by mTALs from sham rats. Apocynin also unmasked a T1D-induced increase in nitrite production. NOS inhibition did not alter superoxide production in either group. In sham mTAL, total NOS inhibition, but not isoform-specific inhibition of NOS1 or NOS2, increased ouabain- and furosemide-sensitive O2 consumption, confirming a tonic inhibitory impact of NOS3 on sodium transport. In contrast, neither total nor isoform-specific NOS inhibition altered O2 consumption by STZ mTAL. Apocynin treatment of STZ mTAL unveiled the ability of isoform-specific NOS inhibition to significantly increase O2 consumption, without further increase in O2 consumption with total NOS inhibition. CONCLUSION: Under normal conditions, NOS3-derived NO inhibits sodium transport in the mTAL. T1D dismantles the impact of NOS-mediated inhibition of sodium transport as a result of NADPH oxidase-dependent NO scavenging. Inhibition of NADPH oxidase to preserve NO bioavailability reveals an inhibitory impact of NOS1- and NOS2-derived NO on sodium transport in the mTAL.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , NADPH Oxidasas/metabolismo , Nefronas/metabolismo , Óxido Nítrico Sintasa/metabolismo , Sodio/metabolismo , Animales , Diabetes Mellitus Tipo 1/metabolismo , Transporte Iónico/fisiología , Masculino , Nefronas/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
AIM: To elucidate the role of the O(2)(-), H(2)O(2) or NO pathways in aortic angiotensin (Ang)II-induced vasoconstriction in Dahl salt-sensitive (SS) rats compared with control SS-13(BN) rats on a normal or hypercaloric diet. METHODS: Aortic function was assessed using wire myography in 16-week-old rats maintained on a normal diet or a 4-week hypercaloric diet. Nitric oxide synthase (NOS) activity and expression was determined by the conversion of radio-labelled arginine to citrulline and Western blot analysis respectively. RESULTS: On normal diet, AngII-induced vasoconstriction was greater in SS than SS-13(BN) rats. Polyethylene glycol superoxide dismutase (PEG-SOD) reduced the aortic AngII response similarly in both strains on normal diet. Catalase blunted, whereas N(ω)-Nitro-L-arginine methyl ester (L-NAME) did not affect the AngII response in SS rats. In SS-13(BN) rats, catalase had no effect and L-NAME enhanced AngII response. On hypercaloric diet, aortic AngII responsiveness was reduced in SS but unaltered in SS-13(BN) rats compared with their normal diet counterparts. PEG-SOD reduced the AngII response in both rats on hypercaloric diet. Catalase treatment did not alter aortic AngII response, while L-NAME increased the response in SS rats on hypercaloric diet. In SS-13(BN) rats on hypercaloric diet, catalase reduced and L-NAME did not alter the AngII response. Furthermore, aortic endothelial-dependent vasorelaxation was increased in SS rats on hypercaloric diet compared with normal diet and aortic NOS3 activity and expression was increased. CONCLUSION: A short-term hypercaloric diet induces a blunted vasoconstrictive and enhanced vasodilatory phenotype in SS rats, but not in SS-13(BN) rats, via reduced H(2)O(2) and increased NOS3 function.
Asunto(s)
Aorta/metabolismo , Dieta/efectos adversos , Óxido Nítrico Sintasa de Tipo III/metabolismo , Vasoconstricción/fisiología , Vasodilatación/fisiología , Animales , Aorta/fisiopatología , Western Blotting , Ingestión de Energía , Ratas , Ratas Endogámicas DahlRESUMEN
AIMS/HYPOTHESIS: Activation of endothelin receptor-A (ET(A)) increases glomerular permeability to albumin (P(alb)) and elevates pro-inflammatory markers in hyperglycaemic rats. METHODS: Male Sprague-Dawley rats were given streptozotocin (n = 32) or saline (sham; n = 32). Half of the animals in each group received the ET(A)-selective antagonist, ABT-627 (atrasentan; orally), beginning immediately after hyperglycaemia was confirmed. Glomeruli were isolated by sieving techniques and P(alb) determined from the change in glomerular volume induced by oncotic gradients of albumin. Glomerular nephrin levels were assessed by immunofluorescence, whereas urinary nephrin was measured by immunoassay. RESULTS: At 3 and 6 weeks after streptozotocin injection, proteinuria was significantly increased compared with sham controls and significantly reduced by ABT-627 treatment. P(alb) was also increased at 3 and 6 weeks post-streptozotocin. ABT-627 had no effect on P(alb) or protein excretion in sham control rats. In glomeruli isolated from hyperglycaemic rats, incubation with BQ-123, a selective ET(A) antagonist, reduced P(alb), whereas BQ-788, a selective endothelin receptor-B antagonist had no effect (n = 6 rats per group, 5-8 glomeruli per rat). Glomerular and plasma content of soluble intercellular adhesion molecule-1 and monocyte chemoattractant protein-1 were significantly increased 6 weeks after streptozotocin (ELISA). ABT-627 attenuated these increases. After 6 weeks of hyperglycaemia, glomerular nephrin content was decreased with a concurrent increase in urinary nephrin excretion. ABT-627 prevented glomerular nephrin loss in hyperglycaemic rats (n = 5-8 rats per group; eight groups). CONCLUSIONS/INTERPRETATION: These observations support the hypothesis that endothelin-1, via the ET(A) receptor, directly increases P(alb), possibly via nephrin loss, as well as early inflammation in the hyperglycaemic rat.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Glomérulos Renales/metabolismo , Receptor de Endotelina A/metabolismo , Animales , Atrasentán , Diabetes Mellitus Experimental/inmunología , Modelos Animales de Enfermedad , Antagonistas de los Receptores de la Endotelina A , Inmunoensayo , Glomérulos Renales/efectos de los fármacos , Glomérulos Renales/inmunología , Masculino , Proteínas de la Membrana/metabolismo , Proteínas de la Membrana/orina , Oligopéptidos/farmacología , Piperidinas/farmacología , Pirrolidinas/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
AIM: Our laboratory and others have shown that endothelin (ET)-1 directly stimulates nitric oxide (NO) production in inner medullary collecting duct (IMCD) cells. The goal of this study was to determine which NO synthase (NOS) isoforms in IMCD are sensitive to ET-1, and the role of ET(A) and ET(B) receptor activation in vivo and in vitro. METHODS: NOS enzymatic activity and NOS isoform protein expression were examined in cultured IMCD-3 cells and isolated renal inner medulla. ET(B) receptor-deficient homozygous rats (sl/sl) have elevated levels of circulating ET-1 and lack a functional ET(B) signalling pathway in kidneys, and furthermore provides a unique model to study ET(A) receptor signalling in the renal inner medulla in vivo. RESULTS: Incubation of IMCD-3 cells with exogenous ET-1 (50 nm) resulted in ET(A)-dependent increased NOS1 protein expression in IMCD-3 cells with no effect on NOS2 or NOS3 expression. ET(B) receptor antagonism has no effect on NOS expression in IMCD-3 cells. Consistent with in vitro results, cytosolic NOS1 protein expression was significantly greater in the renal inner medulla of sl/sl rats compared with heterozygous (sl/+) controls, with no alteration in NOS3 expression. In contrast to protein expression data, NOS1- and NOS3-specific enzymatic activities decreased in the cytosolic fraction from the renal inner medulla of sl/sl compared with sl/+. CONCLUSION: These results provide evidence that both ET(A) and ET(B) receptors regulate NOS isoform activity in the renal inner medulla and specifically support the hypothesis that ET(A) receptor activation increases NOS1 expression.
Asunto(s)
Endotelina-1/metabolismo , Médula Renal/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Óxido Nítrico Sintasa de Tipo I/metabolismo , Receptor de Endotelina A/metabolismo , Receptor de Endotelina B/metabolismo , Sodio/metabolismo , Animales , Células Cultivadas , Femenino , Tasa de Filtración Glomerular/fisiología , Masculino , RatasRESUMEN
Hypertension is associated with increased reactive oxygen species (ROS). Renal ROS production and their effects on renal function have never been investigated in mineralocorticoid hypertensive rats. In this study we hypothesized that increased ROS production in kidneys from deoxycorticosterone (DOCA)-salt rats contributes to adverse renal morphological changes and impaired renal function in DOCA-salt hypertensive rats. We also determined whether ROS-induced renal injury was dependent on blood pressure. DOCA-salt hypertensive rats exhibited a marked increase in blood pressure, renal ROS production, glomerular and tubular lesions, and microalbuminuria compared to sham rats. Treatment of DOCA-salt hypertensive rats with apocynin for 28 days resulted in attenuation of systolic blood pressure and improvement of renal morphology. Renal superoxide level in DOCA-salt rats was 215% of sham-operated rats and it was significantly decreased to 140% with apocynin treatment. Urinary protein level was decreased from 27 +/- 3 mg/day in DOCA-salt hypertensive rats to 9 +/- 2 mg/day. 28 days of Vitamin E treatment also reduced renal injury in regard to urinary protein level and renal morphology but had no effect on blood pressure in DOCA-salt rats. Increased urinary 8-isoprostane, a marker for oxidative stress, in DOCA-salt hypertensive rats (55 +/- 8 ng/day) was diminished by vitamin E treatment (24 +/- 6 ng/day). These data suggest that renal injury characteristic of mineralocorticoid hypertension is associated with oxidative stress and is partly independent of blood pressure.
Asunto(s)
Antioxidantes/farmacología , Hipertensión/fisiopatología , Enfermedades Renales/fisiopatología , Especies Reactivas de Oxígeno/efectos adversos , Especies Reactivas de Oxígeno/metabolismo , Vitamina E/farmacología , Acetofenonas/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Desoxicorticosterona , Histocitoquímica , Hipertensión/inducido químicamente , Enfermedades Renales/tratamiento farmacológico , Glomérulos Renales/patología , Masculino , Proteinuria/prevención & control , Ratas , Ratas Sprague-Dawley , Superóxidos/efectos adversos , Superóxidos/análisisRESUMEN
Endothelium-derived nitric oxide (NO) is a key signalling molecule in the maintenance of cardiovascular health. Endothelial NO synthase (NOS 3), which catalyses the formation of NO, is targeted to the plasma membrane by dual acylation. In vitro studies suggest that membrane localization of NOS 3 is an important regulatory element of NO production. Dysfunction of the vascular endothelium and a decrease in NO bioavailability is associated with the development and progression of a number of cardiovascular diseases, including hypertension. Our laboratory has previously published that in salt-dependent hypertension there is an altered localization of NOS 3, with an increase in cytosolic expression. These data have led us to question whether the increased cytosolic NOS 3 expression is a form of compensation for endothelial dysfunction in hypertension, or an indicator and contributing factor to endothelial dysfunction. This review will outline the importance of subcellular localization in the regulation of NOS 3 in vitro, the role of NOS 3 in endothelial dysfunction associated with salt-dependent hypertension, and the potential physiological consequences of altered NOS 3 localization in vivo.
Asunto(s)
Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico/biosíntesis , Acilación , Animales , Disponibilidad Biológica , Membrana Celular/enzimología , Células Endoteliales/enzimología , Endotelio Vascular/enzimología , Humanos , Hipertensión/metabolismo , Arterias Mesentéricas/enzimología , Óxido Nítrico Sintasa de Tipo III , Ratas , Cloruro de Sodio/metabolismo , Fracciones Subcelulares/enzimologíaRESUMEN
Developmental changes in the lung occur at birth, allowing for the transition from placental to air breathing. Here we have measured nitric oxide synthase (NOS) activity in the porcine lung pre and post partum. NOS activity, which was predominantly calcium dependent, was low in full term fetal tissue compared to that present in lungs from the newborn (5 minutes post partum), 1, 3, 6 and 14 day old animals. No increase in activity was seen when fetal pigs were allowed to breathe for 5 minutes. Specific activity remained low in fetal tissue following partial purification. By contrast, levels of NOS III protein in tissue extracts and in pulmonary arterial endothelial cells, demonstrated by immunohistochemistry, were similar in tissue from the fetal and newborn animals. Thus NOS activity is significantly lower in fetal when compared to postnatal lung tissue despite comparable amounts of NOS III protein being expressed, and birth is followed by an abrupt increase in enzyme activity in animals born at term which correlates with an increase in protein expression.
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Pulmón/enzimología , Pulmón/crecimiento & desarrollo , Óxido Nítrico Sintasa/metabolismo , Envejecimiento , Animales , Animales Recién Nacidos/metabolismo , Western Blotting , Endotelio Vascular/enzimología , Hipertensión Pulmonar/enzimología , Hipertensión Pulmonar/etiología , Hipoxia/complicaciones , Pulmón/embriología , Óxido Nítrico Sintasa/análisis , Óxido Nítrico Sintasa de Tipo II , Óxido Nítrico Sintasa de Tipo III , Respiración , PorcinosRESUMEN
Several disease states, including hypertension, are associated with elevations in plasma endothelin-1 (ET-1) and variable changes in vascular contraction to ET-1. The spotting lethal (sl) rat carries a deletion of the endothelin-B (ET(B)) receptor gene that prevents expression of functional ET(B) receptors, resulting in elevated plasma ET-1. On a normal diet, these rats are normotensive and thus provide an opportunity to study the vascular effects of chronically elevated ET-1 in the absence of hypertension. Studies were performed in rats homozygous for the ET(B) deficiency (sl/sl; n = 8) and in transgenic rats heterozygous for the ET(B) deficiency (sl/+; n = 8). Plasma ET-1 was elevated in sl/sl rats (3.85 +/- 0.55 pg/ml) compared with sl/+ rats (0.31 +/- 0.11 pg/ml). Mean arterial blood pressure in conscious unrestrained sl/sl and sl/+ rats was 101 +/- 5 and 107 +/- 6 mmHg, respectively. Concentration-dependent contractions to ET-1 (10(-11)-10(-8) M) were reduced in mesenteric small arteries (150-250 microm) from sl/sl rats, as indicated by an approximately 10-fold increase in EC(50). A selective ET(A) antagonist, A-127722 (30 nM), abolished contraction to ET-1 in both groups, whereas a selective ET(B) antagonist had no effect. Also, ET(B) agonists (IRL-1620 and sarafatoxin 6c) produced neither contraction nor relaxation in either group, indicating that contraction to ET-1 in this vascular segment was exclusively ET(A) dependent. Despite increased plasma ET-1, protein expression of ET(A) receptors in membrane protein isolated from mesenteric small arteries was increased in sl/sl compared with sl/+ rats, as shown by Western blotting. These results indicate that, in ET(B)-deficient rats, ET(A)-induced contraction is reduced in vessels normally lacking ET(B)-mediated effects. Reduced contraction may be related to elevated plasma ET-1 and occurs in the presence of increased ET(A) receptor protein expression, suggesting an uncoupling of ET(A) receptor expression from functional activity.
Asunto(s)
Endotelina-1/farmacología , Receptores de Endotelina/genética , Receptores de Endotelina/metabolismo , Vasoconstricción/efectos de los fármacos , Vasoconstricción/fisiología , Animales , Animales Modificados Genéticamente , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Western Blotting , Relación Dosis-Respuesta a Droga , Endotelina-1/sangre , Heterocigoto , Homocigoto , Masculino , Arterias Mesentéricas/química , Arterias Mesentéricas/fisiología , Ratas , Receptor de Endotelina A , Receptor de Endotelina B , Receptores de Endotelina/análisis , Resistencia Vascular/efectos de los fármacos , Resistencia Vascular/fisiologíaRESUMEN
BACKGROUND: The relationship between urinary vasoactive factors and sodium excretion has not been adequately addressed in humans. PROCEDURE: Excretion rates of sodium, nitrates/nitrites (NOx), cGMP, and endothelin-1 (ET-1) were measured before and after ingestion of a mixed electrolyte solution (8 oz Gatorade) while undergoing a routine cardiovascular evaluation in a sample of 51 normotensive young adults. RESULTS: Significant correlations were detected for changes in excretion between all four variables, r ranged from 0.50 to 0.86 (P < .001). Correlations were higher in African Americans than white Americans. CONCLUSIONS: The association of renal ET-1 and NO activity with sodium excretion supports the hypothesis that these factors play a role in the physiologic response to acute changes in sodium intake, particularly in African Americans.
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GMP Cíclico/orina , Endotelina-1/orina , Óxido Nítrico/orina , Sodio/orina , Adolescente , Adulto , Población Negra , GMP Cíclico/fisiología , Endotelina-1/fisiología , Femenino , Humanos , Hipertensión/etiología , Masculino , Natriuresis , Óxido Nítrico/fisiología , Estados Unidos , Población BlancaRESUMEN
The aim of this study was to determine the response of inflammatory and vasoactive mediators to 3 consecutive days of exercise in African-American women with and without sickle cell anemia (SCA). Circulating inflammatory mediators [C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha)] were measured before, and vasoactive mediators [endothelin-1 (ET-1), nitric oxide metabolites (NOx)] before and after each exercise bout in ten subjects with SCA and ten controls. Exercise did not affect ET-1, IL-6 or CRP concentrations (p >.05). TNFalpha was higher in SCA than controls (p < or = .0005) at all times; however, the response pattern was similar for the groups: no change from day 1 to day 2, but a decrease from day 2 to day 3 (p < or = .05). NOx increased significantly after exercise (p < or = .0001) but returned to baseline by 24 h afterward. On the 3rd day, NOx increased after exercise in SCA but not in the controls (p < or = .05). In conclusion, exercise did not cause a harmful inflammatory response in these individuals with SCA. However, NOx increased after exercise on all 3 days in SCA but appeared attenuated after 2 days in controls.
Asunto(s)
Anemia de Células Falciformes/sangre , Ejercicio Físico , Adulto , Anemia de Células Falciformes/diagnóstico , Anemia de Células Falciformes/fisiopatología , Biomarcadores/sangre , Proteína C-Reactiva/análisis , Endotelina-1/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Polarización de Fluorescencia , Frecuencia Cardíaca , Hematócrito , Hemoglobinas/análisis , Humanos , Inmunoensayo , Interleucina-6/sangre , Óxido Nítrico/sangre , Dolor/diagnóstico , Factores de Tiempo , Factor de Necrosis Tumoral alfa/análisisRESUMEN
Recent evidence suggests that endothelin-1 (ET-1), perhaps through the ET(B) receptor, may participate in blood pressure regulation through the control of sodium excretion. Mean arterial pressure (MAP) was continuously measured via telemetry implants in male Sprague-Dawley rats. After 1 wk of baseline measurements, rats were given either high (10%) or low (0.08%) NaCl in chow for the remainder of the experiment (n = 5 in each group). MAP was significantly increased in rats on a high-salt diet (115 +/- 2 mmHg) compared with rats on the low-salt diet (103 +/- 2 mmHg; P < 0.05). All rats were then treated with the ET(B) receptor antagonist A-192621 mixed with the food and adjusted daily to ensure a dose of 30 mg x kg(-1) x day(-1). ET(B) blockade produced an increase in MAP within a few hours of treatment and was significantly higher in rats on the high-salt diet over a 1-wk period (170 +/- 3 vs. 115 +/- 3 mmHg, P < 0.01). To determine whether the increase in MAP during A-192621 treatment was due to increased ET(A) receptor activation, all rats were then given the ET(A)-selective antagonist ABT-627 in the drinking water while a low-salt/high-salt diet and ET(B) blockade were continued. ABT-627 decreased MAP within a few hours in rats on either the high-salt (113 +/- 3 mmHg) or low-salt (101 +/- 3 mmHg) diet. These results support the hypothesis that endothelin, through the ET(B) receptor, participates in blood pressure regulation in the response to salt loading.
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Antagonistas de los Receptores de Endotelina , Endotelinas/metabolismo , Pirrolidinas/farmacología , Sodio en la Dieta/administración & dosificación , Animales , Presión Sanguínea/efectos de los fármacos , Endotelinas/orina , Frecuencia Cardíaca/efectos de los fármacos , Ratas , Receptor de Endotelina B , Cloruro de Sodio/administración & dosificación , Telemetría , Factores de TiempoRESUMEN
OBJECTIVE: The goal of this work was to test the functional role of vascular endothelial growth factor (VEGF) in promoting the vigorous granulation tissue formation, wound fluid accumulation, and angiogenic responses characteristic of this wound model. BACKGROUND: Formation of vessel-rich granulation tissue is central to wound repair and is thought to be regulated by locally liberated angiogenic factors. Despite the clinical importance of granulation tissue formation in the early stage of wound healing, surprisingly little is known about the molecular identity of signals leading to granulation tissue invasion of a wound space. Methods. A ventral hernia, surgically created in the abdominal wall of 15 swine, was repaired using silicone sheeting and skin closure. An osmotic minipump, inserted in a remote subcutaneous pocket, delivered saline (n = 5), an irrelevant control antibody (n = 5), or neutralizing anti-VEGF antibody (n = 5) into the wound environment. Serial ultrasonography on Days 2, 4, 7, 9, 11, and 14 was used to determine the dimensions of the subcutaneous granulation tissue and wound fluid compartment. VEGF and transforming growth factor beta1 (TGF-beta1) levels in serial wound fluid samples were quantitated by ELISA. On Day 14, animals were sacrificed and the abdominal wall was harvested for histologic, biochemical, and molecular analyses. RESULTS: In animals receiving saline or an irrelevant antibody, a nearly linear 4-fold increase in granulation tissue thickness and 7-fold increase in wound fluid volume were measured over the 14-day study interval. In contrast, in animals receiving anti-VEGF neutralizing antibody, Day 14 granulation tissue thickness and wound fluid volume measurements were essentially unchanged from Day 2 values. Moreover, in the anti-VEGF animals, ultrasonography was unable to resolve the "angiogenic zone" typical of both controls, and correspondingly, wound vessel count and vascular surface area estimates derived from image analysis of histological sections were 3-fold lower in the anti-VEGF animals compared with the saline and antibody controls. Finally, VEGF levels in wound fluid detectable by ELISA analysis were strikingly (10-fold) reduced in anti-VEGF animals on Postsurgery Days 7-14. In contrast, TGF-beta1 levels were unaffected by the anti-VEGF treatment. CONCLUSION: Functional VEGF is a key mediator in wound angiogenesis, fluid accumulation, and granulation tissue formation.
Asunto(s)
Factores de Crecimiento Endotelial/fisiología , Tejido de Granulación/crecimiento & desarrollo , Linfocinas/fisiología , Cicatrización de Heridas/fisiología , Músculos Abdominales/lesiones , Animales , Anticuerpos/administración & dosificación , Anticuerpos/inmunología , Anticuerpos/farmacología , Especificidad de Anticuerpos , Líquidos Corporales/efectos de los fármacos , Líquidos Corporales/metabolismo , Factores de Crecimiento Endotelial/inmunología , Femenino , Tejido de Granulación/efectos de los fármacos , Hernia Ventral/patología , Hernia Ventral/fisiopatología , Linfocinas/inmunología , Neovascularización Fisiológica/efectos de los fármacos , Neovascularización Fisiológica/fisiología , Porcinos , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial Vascular , Cicatrización de Heridas/efectos de los fármacos , Heridas y Lesiones/patología , Heridas y Lesiones/fisiopatologíaRESUMEN
BACKGROUND: This study was designed to determine whether the nitric oxide (NO) pathway is involved in wound granulation tissue formation. METHODS: A section of the pig abdominal wall (excluding the skin) was excised, creating an incisional hernia. The resulting defect was repaired with silicone sheeting in a manner that mimics a temporary abdominal wall closure. During the 14-day experimental period, porcine omentum adhered to the peritoneal edges of the defect and a highly vascularized granulation tissue formed on both sides of the sheeting. Granulation tissue thickness and wound fluid volume were monitored by ultrasonography and epigastric artery flow velocity was monitored by color Doppler flow analysis at days 2, 4, 7, 9, 11, and 14. Fluid was serially harvested from the wound compartment at days 2, 4, 7, 9, 11, and 14 for nitrite/ nitrate (NOx) analysis. Finally, granulation tissue was harvested at day 14 for immunohistochemical and molecular analyses. RESULTS: There was a significant increase in granulation tissue thickness and wound fluid volume during the 14-day study period. Blood flow to the wound increased significantly by day 4 and returned toward baseline by day 14. Wound fluid NOx levels significantly increased from days 7 to 11 and then decreased to near baseline values by day 14. Wound fluid arginine levels significantly decreased when compared with peritoneal fluid and plasma levels at day 14, while wound fluid ornithine levels significantly increased. Immunohistochemical analysis of granulation tissue at day 14 revealed nitric oxide synthase (NOS) 2 was present in the majority of the cells in the granulation tissue. NOS 3 was expressed in endothelial cells only, and NOS 1 expression was not observed in the granulation tissue. CONCLUSIONS: This study suggests that NO, NOS 2, and arginine may play critical roles in granulation tissue formation and wound healing. Arginase and NOS 2 may compete for available arginine as a substrate, thereby limiting later NO production in favor of sustained ornithine synthesis.
Asunto(s)
Tejido de Granulación/enzimología , Óxido Nítrico Sintasa/metabolismo , Traumatismos Abdominales/diagnóstico por imagen , Traumatismos Abdominales/enzimología , Traumatismos Abdominales/fisiopatología , Músculos Abdominales/irrigación sanguínea , Animales , Arterias/diagnóstico por imagen , Líquidos Corporales/metabolismo , Tejido de Granulación/diagnóstico por imagen , Inmunohistoquímica , Nitratos/metabolismo , Óxido Nítrico Sintasa de Tipo I , Óxido Nítrico Sintasa de Tipo II , Óxido Nítrico Sintasa de Tipo III , Nitritos/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Porcinos , Ultrasonografía Doppler en Color , Cicatrización de HeridasRESUMEN
Hyperinsulinemia, a primary feature of insulin resistance, is associated with increased endothelin-1 (ET-1) activity. This study determined the vascular response to ET-1 and receptor binding characteristics in small mesenteric arteries of insulin-resistant (IR) rats. Rats were randomized to control (C) (n = 32) or IR (n = 32) groups. The response to ET-1 was assessed (in vitro) in arteries with (Endo+) and without (Endo-) endothelium. In addition, arteries (Endo+) were pretreated with the ET(B) antagonist A-192621 or the ET(A) antagonist A-127722. Finally, binding characteristics of [(125)I]ET-1 were determined. Results showed that in Endo+ arteries the maximal relaxation (E(max)) to ET-1 was similar between C and IR groups; however, the concentration at 50% of maximum relaxation (EC(50)) was decreased in IR arteries. In Endo- arteries, the E(max) to ET-1 was enhanced in both groups. Pretreatment with A-192621 enhanced the E(max) and EC(50) to ET-1 in both groups. In contrast, A-127722 inhibited the ET-1 response in all arteries in a concentration-dependent manner; however, a greater ET-1 response was seen at each concentration in IR arteries. Maximal binding of [(125)I]ET-1 was increased in IR versus C arteries although the dissociation constant values were similar. In conclusion, we found the vasoconstrictor response to ET-1 is enhanced in IR arteries due to an enhanced expression of ET receptors and underlying endothelial dysfunction.
Asunto(s)
Endotelina-1/metabolismo , Hiperinsulinismo/metabolismo , Resistencia a la Insulina/fisiología , Arterias Mesentéricas/metabolismo , Receptores de Endotelina/metabolismo , Animales , Peso Corporal , Relación Dosis-Respuesta a Droga , Antagonistas de los Receptores de Endotelina , Endotelina-1/biosíntesis , Técnicas In Vitro , Radioisótopos de Yodo , Masculino , Arterias Mesentéricas/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/metabolismo , Pirrolidinas/farmacología , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-Dawley , Receptor de Endotelina A , Receptor de Endotelina B , Receptores de Endotelina/biosíntesisRESUMEN
Exercise training produces enhanced nitric oxide (NO)-dependent, endothelium-mediated vasodilator responses of porcine coronary arterioles but not conduit coronary arteries. The purpose of this study was to test the hypothesis that exercise training increases the amount of endothelial NO synthase (eNOS) in the coronary arterial microcirculation but not in the conduit coronary arteries. Miniature swine were either exercise trained or remained sedentary for 16--20 wk. Exercise-trained pigs exhibited increased skeletal muscle oxidative capacity, exercise tolerance, and heart weight-to-body weight ratios. Content of eNOS protein was determined with immunoblot analysis in conduit coronary arteries (2- to 3-mm ID), small arteries (301- to 1,000-microm ID), resistance arteries (151- to 300-microm ID), and three sizes of coronary arterioles [large (101- to 150-microm ID), intermediate (51- to 100-microm ID), and small (<50-microm ID)]. Immunoblots revealed increased eNOS protein in some sizes of coronary arteries and arterioles but not in others. Content of eNOS was increased by 60--80% in small and large arterioles, resistance arteries, and small arteries; was increased by 10--20% in intermediate-sized arterioles; and was not changed or decreased in conduit arteries. Immunohistochemistry revealed that eNOS was located in the endothelial cells in all sizes of coronary artery. We conclude that exercise training increases eNOS protein expression in a nonuniform manner throughout the coronary arterial tree. Regional differences in shear stress and intraluminal pressures during exercise training bouts may be responsible for the distribution of increased eNOS protein content in the coronary arterial tree.
Asunto(s)
Vasos Coronarios/enzimología , Endotelio Vascular/enzimología , Óxido Nítrico Sintasa/biosíntesis , Esfuerzo Físico , Animales , Arterias/anatomía & histología , Arterias/enzimología , Arteriolas/anatomía & histología , Arteriolas/enzimología , Vasos Coronarios/anatomía & histología , Prueba de Esfuerzo , Tolerancia al Ejercicio , Femenino , Immunoblotting/métodos , Inmunohistoquímica , Óxido Nítrico Sintasa/inmunología , PorcinosRESUMEN
Recent evidence suggests that nitric oxide (NO) within the inner medullary collecting duct (IMCD) functions to regulate sodium and water reabsorption. Because fluid shear stress has been shown to increase NO production in endothelial and vascular smooth muscle cells, experiments were designed to determine whether a similar mechanism exists in IMCD cells. Cultured IMCD-3 cells derived from murine IMCD were subjected to 60 min of pulsatile shear stress. Nitrite production (2,3-diaminonaphthalene fluorometric assay) increased 12-, 16-, and 23-fold at 3.3, 10, and 30 dyn/cm(2), respectively, compared with static control cultures. Preincubation with the non-isoform-specific NO synthase inhibitor nitro-L-arginine methyl ester reduced nitrite production by 83% in response to 30 dyn/cm(2). Western blotting and immunofluorescence analysis of static IMCD-3 cell cultures revealed the expression of all three NO synthase isoforms (NOS-1 or neuronal NOS, NOS-2 or inducible NOS, and NOS-3 or endothelial NOS) in IMCD-3 cultures. These results indicate that NO production is modulated by shear stress in IMCD-3 cells and that fluid shear stress within the renal tubular system may play a role in the regulation of sodium and water excretion by control of NO production in the IMCD.