RESUMEN
Large deletions in the first intron of the With No lysine (K) 1 (WNK1) gene are responsible for Familial Hyperkalemic Hypertension (FHHt), a rare form of human hypertension associated with hyperkalemia and hyperchloremic metabolic acidosis. We generated a mouse model of WNK1-associated FHHt to explore the consequences of this intronic deletion. WNK1(+/FHHt) mice display all clinical and biological signs of FHHt. This phenotype results from increased expression of long WNK1 (L-WNK1), the ubiquitous kinase isoform of WNK1, in the distal convoluted tubule, which in turn, stimulates the activity of the Na-Cl cotransporter. We also show that the activity of the epithelial sodium channel is not altered in FHHt mice, suggesting that other mechanisms are responsible for the hyperkalemia and acidosis in this model. Finally, we observe a decreased expression of the renal outer medullary potassium channel in the late distal convoluted tubule of WNK1(+/FHHt) mice, which could contribute to the hyperkalemia. In summary, our study provides insights into the in vivo mechanisms underlying the pathogenesis of WNK1-mediated FHHt and further corroborates the importance of WNK1 in ion homeostasis and blood pressure.
Asunto(s)
Túbulos Renales Distales/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Seudohipoaldosteronismo/genética , Animales , Canales Epiteliales de Sodio/metabolismo , Eliminación de Gen , Ratones , Ratones Transgénicos , Antígenos de Histocompatibilidad Menor , Canales de Potasio de Rectificación Interna/genética , Seudohipoaldosteronismo/metabolismo , Proteína Quinasa Deficiente en Lisina WNK 1RESUMEN
OBJECTIVES: Leaflet injury has been documented to occur during the deployment of valved stents (VSs). The pathological aspects, however, of this injury are difficult to quantify. Conversely, the hydraulic conductance of a (pericardial) membrane may be easily determined. The impact of crimping and deployment of VS on this parameter was therefore investigated. METHODS: Bovine pericardial square (25 × 25 mm) patches were placed within a pressure chamber and their hydraulic conductance was determined. The influence of the pressure gradient and tissue thickness on this parameter was analysed. Six balloon-expandable VS were constructed. The hydraulic conductance of their bovine pericardial leaflets was determined before and after VS crimping and deployment in four of them. Pericardial leaflets of non-crimped VS were used as controls. RESULTS: Hydraulic conductance increased insignificantly with the pressure level within the chamber: from 128 ± 26.9 ml/h/m(2)/mmHg at a pressure of 50 mmHg to 232.3 ± 51.9 ml/h/m(2)/mmHg at a pressure of 250 mmHg (P = 0.117). Hydraulic conductance was not correlated to pericardial thickness, for thickness measurements ranging from 0.34 to 0.76 mm. The hydraulic conductance of VS leaflets significantly increased immediately after crimping from 45.2 ± 7.6 to 667.9.0 ± 527.2 ml/h/m(2)/mmHg (P < 0.001). This increase was still observed 24 h after VS deployment. No change in hydraulic conductance occurred in the control group. CONCLUSIONS: The determination of the hydraulic conductance of pericardial patches was easy to perform, reproducible and not influenced by tissue thickness. The hydraulic conductance of pericardial leaflets dramatically increased after VS crimping and deployment. This parameter might be, in the future, a useful noninvasive tool in studying leaflet trauma.
Asunto(s)
Pericardio/fisiología , Diseño de Prótesis , Stents , Animales , Fenómenos Bioquímicos , Bovinos , Ensayo de Materiales , Presión , ReologíaRESUMEN
Since android overweight/obesity and insulin resistance are independent risk factors for cardiovascular disease, we investigated their impact on basal and postprandial scavenger receptor BI (SR-BI) and ATP binding cassette transporter A1 (ABCA1)-mediated serum cholesterol efflux. Twelve android overweight to obese and 9 normal weight controls women underwent body composition analysis by dual energy X-ray absorptiometry, a euglycemic hyperinsulinemic clamp, and an oral fat load with blood sampling at initial time (T0), 4h (T4) and 10h (T10) after the fat load. Serum lipids and HDL-parameters, capacities of serum to promote cholesterol efflux from SR-BI expressing Fu5AH hepatoma cells or from ABCA1-expressing J774 macrophages and to abilities of serum to induce a net removal of cholesterol from macrophage foam cells were measured at T0, T4 and T10. Sera from overweight/obese exhibited moderately decreased SR-BI-mediated cholesterol efflux capacities, in accordance with reduced HDL concentrations, but importantly increased ABCA1-mediated cholesterol efflux and increased cholesterol extraction capacities over the postprandial period, partly related to higher prebeta-HDL concentrations. In multiple regression analyses, android obesity-related parameters and HDL-PL or prebeta-HDL levels remained the only independent correlates for SR-BI or ABCA1-dependent fractional cholesterol efflux while only prebeta-HDL levels remained correlated to cholesterol extraction capacities. Our results suggest that android overweight/obesity may not result in an impaired cholesterol efflux capacity.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/sangre , Colesterol/sangre , Resistencia a la Insulina/fisiología , Obesidad/metabolismo , Sobrepeso/sangre , Receptores Depuradores de Clase B/sangre , Transportador 1 de Casete de Unión a ATP , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 1 , Transportadoras de Casetes de Unión a ATP/biosíntesis , Adulto , Animales , Composición Corporal , Carcinoma Hepatocelular/metabolismo , Línea Celular , Femenino , Células Espumosas/metabolismo , Lipoproteínas de Alta Densidad Pre-beta/fisiología , Humanos , Neoplasias Hepáticas/metabolismo , Macrófagos/metabolismo , Ratones , Persona de Mediana Edad , Periodo Posprandial , Ratas , Células Tumorales Cultivadas , Circunferencia de la CinturaRESUMEN
1. Plasma levels of matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) are potential markers of many diseases involving extracellular matrix remodelling such as hypertension. Our aim was to determine whether the anticoagulant used to collect plasma and several freeze-thaw cycles may influence the accuracy of plasma MMP and TIMP determinations. 2. Plasma samples of 18 healthy volunteers were collected on three anticoagulants: heparinate, citrate and EDTA. For each anticoagulant, we compared: (i) MMP-2 and MMP-9 levels using gelatin zymography and TIMP-1 and TIMP-2 concentrations using enzyme-linked immunosorbent assay; (ii) intra- and interassay coefficients of variation (CV); and (iii) MMP and TIMP levels after up to five freeze-thaw cycles. 3. The choice of anticoagulant influenced TIMP-2 and TIMP-1 concentrations (TIMP-2, P < 0.0001; paired comparisons, citrate vs EDTA, P < 0.0001; EDTA vs heparin, P < 0.0001; citrate vs heparin, P < 0.0001; TIMP-1, P < 0.001; paired comparisons, citrate vs EDTA, P = 0.10; EDTA vs heparin, P < 0.01; citrate vs heparin, P < 0.0001), but not those of MMP. We observed a bias with heparinate for TIMP-2, TIMP-1 and MMP-9 determinations. The anticoagulant did not influence intra-assay or interassay CV. Performing freeze-thaw cycles led to alterations in the TIMP-1 plasma levels (P < 0.0001), regardless of the anticoagulant used, whereas MMP and TIMP-2 concentrations were not significantly affected. 4. Anticoagulant influences the measured levels of MMP and TIMP in plasma and should be systematically reported. However, it does not influence the reproducibility of the measurements. Repeated freeze-thaw cycles alter the measurement of TIMP-1 levels and should be avoided.
Asunto(s)
Recolección de Muestras de Sangre/métodos , Metaloproteinasas de la Matriz/sangre , Inhibidores Tisulares de Metaloproteinasas/sangre , Adulto , Anticoagulantes/química , Femenino , Congelación , Humanos , Masculino , Metaloproteinasas de la Matriz/metabolismo , Persona de Mediana Edad , Inhibidores Tisulares de Metaloproteinasas/metabolismoRESUMEN
Hypertriglyceridemia being an independent cardiovascular risk factor, we have compared the potential of sera from asymptomatic hypertriglyceridemic (HTG) type IIb, type IV or normolipidemic (NLP) subjects to promote both fractional cholesterol efflux and cellular cholesterol mass changes using macrophage foam cells. The J774 cells loaded with cholesterol by incubation with acetylated LDL were incubated in the absence or presence of cAMP to upregulate ABCA1 (ATP binding cassette transporter A1) and then incubated for 24h with 1% serum. Compared with NLP, type IV sera exhibited a major increase in ABCA1-dependent efflux while type IIb sera exhibited a moderate but not significant increased ABCA1-mediated efflux. Moreover, positive correlations were established between ABCA1-dependent efflux and the serum prebeta-HDL or TG concentrations. The major finding was that the sera from type IV induced higher total cholesterol and cholesteryl ester mass depletions from ABCA1-expressing cells compared with other groups. Moreover, negative correlations were obtained between total cholesterol or cholesteryl ester mass changes and serum prebeta-HDL levels. In conclusion, we demonstrated for the first time that the serum prebeta-HDL present in high proportions in type IV HTG subjects are not only responsible for higher cholesterol efflux potential but also for increased abilities to promote net removal of cholesterol from macrophage foam cells.
Asunto(s)
Células Espumosas/metabolismo , Hiperlipoproteinemia Tipo IV/inmunología , Hiperlipoproteinemia Tipo IV/metabolismo , Lipoproteínas LDL/farmacocinética , Transportador 1 de Casete de Unión a ATP , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 1 , Transportadoras de Casetes de Unión a ATP/metabolismo , Adulto , Animales , Apolipoproteína A-I/sangre , Apolipoproteínas B/sangre , Células Cultivadas , HDL-Colesterol/sangre , LDL-Colesterol/sangre , Células Espumosas/citología , Humanos , Masculino , Ratones , Persona de Mediana Edad , Receptores Depuradores de Clase B/metabolismo , Triglicéridos/sangre , TritioRESUMEN
BACKGROUND: Pulmonary expression of heme oxygenase has been observed in multiple studies. This expression has been found beneficial in decreasing the severity of acute lung injury (ALI) post ischemia-reperfusion (I/R). The aim of this study was to assess the role of exogenous administration of the end-products of heme oxygenase reaction, carbon monoxide, and bilirubin, in the severity of ALI. STUDY DESIGN: We compared five groups of rats (n = 7/group) including a sham group and four I/R of the lower extremities by clamping the abdominal aorta for 2 h followed by reperfusion for 2 h. The four I/R groups included a control group, one pretreated with bilirubin (50 micromol/kg IV), another with inhaled carbon monoxide (CO) (250 ppm), and the last pretreated with both. The severity of ALI has been evaluated by a histological assay grading neutrophilic infiltration, as well as a study of the microvascular permeability using the Evans blue. RESULTS: The administration of CO prevented pulmonary microvascular permeability alteration noted after I/R of the lower limbs (pulmonary content of Evans blue: 141 +/- 23 microg/g of tissue in the isolated I/R group versus 68 +/- 34 microg/g of tissue in CO group; P < 0.001). Histologically CO administration inhibited neutrophilic sequestration observed after I/R. On the other hand, treatment by bilirubin alone (50 micromol/kg IV) did not modify the extent of pulmonary injury. CONCLUSION: Exogenous administration of carbon monoxide by inhalation at low doses prevented ALI post-I/R in this model.
Asunto(s)
Antimetabolitos/farmacología , Monóxido de Carbono/farmacología , Daño por Reperfusión/complicaciones , Síndrome de Dificultad Respiratoria/etiología , Síndrome de Dificultad Respiratoria/prevención & control , Animales , Antiinflamatorios/farmacología , Bilirrubina/farmacología , Presión Sanguínea , Colorantes/farmacocinética , Modelos Animales de Enfermedad , Quimioterapia Combinada , Azul de Evans/farmacocinética , Hemo/metabolismo , Miembro Posterior , Masculino , Edema Pulmonar/etiología , Edema Pulmonar/patología , Edema Pulmonar/prevención & control , Ratas , Ratas Wistar , Síndrome de Dificultad Respiratoria/patología , Índice de Severidad de la EnfermedadAsunto(s)
Metaloproteasas/sangre , Inhibidores Tisulares de Metaloproteinasas/sangre , Factores de Edad , Etnicidad , Humanos , Metaloproteinasa 2 de la Matriz/sangre , Metaloproteinasa 9 de la Matriz/sangre , Estándares de Referencia , Factores de Tiempo , Inhibidor Tisular de Metaloproteinasa-1/sangre , Inhibidor Tisular de Metaloproteinasa-2/sangreRESUMEN
NH(4)(+) transport by the distal nephron and NH(4)(+) detoxification by the liver are critical for achieving regulation of acid-base balance and to avoid hyperammonemic hepatic encephalopathy, respectively. Therefore, it has been proposed that rhesus type B glycoprotein (Rhbg), a member of the Mep/Amt/Rh NH(3) channel superfamily, may be involved in some forms of distal tubular acidosis and congenital hyperammonemia. We have tested this hypothesis by inactivating the RHbg gene in the mouse by insertional mutagenesis. Histochemical studies analyses confirmed that RHbg knockout (KO) mice did not express Rhbg protein. Under basal conditions, the KO mice did not exhibit encephalopathy and survived well. They did not exhibit hallmarks of distal tubular acidosis because neither acid-base status, serum potassium concentration, nor bone mineral density was altered by RHbg disruption. They did not have hyperammonemia or disturbed hepatic NH(3) metabolism. Moreover, the KO mice adapted to a chronic acid-loading challenge by increasing urinary NH(4)(+) excretion as well as their wild-type controls. Finally, transepithelial NH(3) diffusive permeability, or NH(3) and NH(4)(+) entry across the basolateral membrane of cortical collecting duct cells, measured by in vitro microperfusion of collecting duct from KO and wild-type mice, was identical with no apparent effect of the absence of Rhbg protein. We conclude that Rhbg is not a critical determinant of NH(4)(+) excretion by the kidney and of NH(4)(+) detoxification by the liver in vivo.
Asunto(s)
Amoníaco/metabolismo , Glicoproteínas/genética , Riñón/fisiología , Proteínas de Transporte de Membrana/genética , Acidosis Tubular Renal/fisiopatología , Amoníaco/orina , Animales , Proteína 1 de Intercambio de Anión de Eritrocito/metabolismo , Densidad Ósea , Ratones , Ratones Noqueados , Mutagénesis Insercional , ATPasas de Translocación de Protón/biosíntesisRESUMEN
The Cl(-)/HCO(3)(-) exchanger AE2 is believed to be involved in transcellular bicarbonate reabsorption that occurs in the thick ascending limb of Henle's loop (TAL). The purpose of this study was to test whether chronic changes in acid-base status and sodium intake regulate AE2 polypeptide abundance in the TAL of the rat. Rats were subjected to 6 d of loading with NaCl, NH(4)Cl, NaHCO(3), KCl, or KHCO(3). AE2 protein abundance was estimated by semiquantitative immunoblotting in renal membrane fractions isolated from the cortex and the outer medulla of treated and control rats. In the renal cortex, AE2 abundance was markedly increased in response to oral loading with NH(4)Cl or with NaCl. In contrast, AE2 abundance was unchanged in response to loading with KCl or with NaHCO(3) and was decreased by loading with KHCO(3). The response of AE2 in the outer medulla differed from that in the cortex in that HCO(3)(-) loading increased AE2 abundance when administered with Na(+) but had no effect when administered with K(+). Immunohistochemistry revealed that NaCl loading increased AE2 abundance in the basolateral membrane of both the cortical and the medullary TAL. In contrast, NH(4)Cl loading increased AE2 abundance only in the cortical TAL but not in the medullary TAL. These results suggest that regulation of the basolateral Cl(-)/HCO(3)(-) exchanger AE2 plays an important role in the adaptation of bicarbonate absorption in the TAL during chronic acid-base disturbances and high sodium intake. The present study also emphasizes the contribution of cortical TAL adaptation in the renal regulation of acid-base status.
Asunto(s)
Equilibrio Ácido-Base/fisiología , Acidosis/metabolismo , Proteínas de Transporte de Anión/metabolismo , Antiportadores/metabolismo , Asa de la Nefrona/metabolismo , Sodio/metabolismo , Equilibrio Ácido-Base/efectos de los fármacos , Administración Oral , Alcalosis/metabolismo , Cloruro de Amonio/farmacología , Animales , Bicarbonatos/farmacología , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/fisiología , Inmunohistoquímica , Corteza Renal/metabolismo , Médula Renal/metabolismo , Asa de la Nefrona/efectos de los fármacos , Masculino , Cloruro de Potasio/farmacología , Compuestos de Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Proteínas SLC4A , Bicarbonato de Sodio/farmacología , Cloruro de Sodio/farmacologíaRESUMEN
Pendrin (Pds; Slc26A4) is a new anion exchanger that is believed to mediate apical Cl(-)/HCO(3)(-) exchange in type B and non-A-non-B intercalated cells of the connecting tubule and cortical collecting duct. Recently, it has been proposed that this transporter may be involved in NaCl balance and blood pressure regulation in addition to its participation in the regulation of acid-base status. The purpose of our study was to determine the regulation of Pds protein abundance during chronic changes in chloride balance. Rats were subjected to either NaCl, NH(4)Cl, NaHCO(3), KCl, or KHCO(3) loading for 6 days or to a low-NaCl diet or chronic furosemide administration. Pds protein abundance was estimated by semiquantitative immunoblotting in renal membrane fractions isolated from the cortex of treated and control rats. We observed a consistent inverse relationship between Pds expression and diet-induced changes in chloride excretion independent of the administered cation. Conversely, NaCl depletion induced by furosemide was associated with increased Pds expression. We conclude that Pds expression is specifically regulated in response to changes in chloride balance.
Asunto(s)
Antiportadores de Cloruro-Bicarbonato/análisis , Cloruros/metabolismo , Homeostasis/fisiología , Riñón/química , Cloruro de Amonio/administración & dosificación , Animales , Bicarbonatos/administración & dosificación , Membrana Celular/química , Cloruros/administración & dosificación , Dieta , Furosemida/administración & dosificación , Homeostasis/efectos de los fármacos , Immunoblotting , Masculino , Cloruro de Potasio/administración & dosificación , Compuestos de Potasio/administración & dosificación , Ratas , Ratas Sprague-Dawley , Bicarbonato de Sodio/administración & dosificación , Cloruro de Sodio/administración & dosificación , Cloruro de Sodio Dietético/administración & dosificación , Transportadores de SulfatoRESUMEN
OBJECTIVE: Expression of inducible heme oxygenase has been shown to be increased in various visceral inflammatory disorders, which may confer a protective role. The purpose of our study was to determine whether the expression of inducible heme oxygenase was up-regulated within lungs in a rat model of extracorporeal circulation. METHODS: Wistar rats underwent either a partial femoro-femoral extracorporeal circulation in normothermia for 3 hours (n = 5) or a sham procedure (n = 5). Exhaled carbon monoxide concentration was monitored with an infrared analyzer. After the rats were killed, lungs were harvested for determination of heme oxygenase activity and inducible heme oxygenase expression (by Western blot and immunohistochemistry). Lung injury was also assessed by arterial blood gas analysis and microscopic study. RESULTS: Extracorporeal circulation was responsible for a lung injury characterized by decreased arterial blood oxygen saturation and typical morphologic findings (marked alveolar neutrophil infiltration; interstitial edema). Exhaled carbon monoxide concentration remained stable throughout the experiment in all sham rats, whereas it increased after extracorporeal circulation (from 0.16 +/- 0.05 ppm at baseline to 0.7 +/- 0.2 ppm at end of experiment; P =.0001). Pulmonary heme oxygenase activity and inducible heme oxygenase content (assessed by Western blot) were increased within lungs of rats that underwent extracorporeal circulation. Immunohistochemistry revealed that the expression of inducible heme oxygenase was mainly localized to inflammatory cells. CONCLUSIONS: Post-extracorporeal circulation acute lung injury in rats was associated with an increased expression of inducible heme oxygenase, the functional significance of which remains to be determined.
Asunto(s)
Pruebas Respiratorias , Monóxido de Carbono/análisis , Hemo Oxigenasa (Desciclizante)/biosíntesis , Daño por Reperfusión/metabolismo , Síndrome de Dificultad Respiratoria/metabolismo , Enfermedad Aguda , Animales , Western Blotting , Circulación Extracorporea , Hemo-Oxigenasa 1 , Inmunohistoquímica , Pulmón/metabolismo , Pulmón/patología , Masculino , Ratas , Ratas Wistar , Daño por Reperfusión/etiología , Síndrome de Dificultad Respiratoria/etiología , Síndrome de Dificultad Respiratoria/patología , Regulación hacia ArribaRESUMEN
Since elevated plasma triglycerides (TGs) are an independent cardiovascular risk factor, we have compared the cholesterol efflux potential of sera from asymptomatic hypertriglyceridemic (HTG) type IIb, type IV or normolipidemic (NLP) individuals using two different cell systems. In both type IIb and IV HTG, the efflux of cholesterol from SR-BI-rich Fu5AH cells was similar to that obtained with NLP. The maintenance of efflux efficiency in spite of reduced HDL-cholesterol levels can be mainly attributed to the relative enrichment of HDL with phospholipid. In the J774 macrophage cell system, pretreatment with cAMP, which upregulates ABCA1, induced a markedly higher increase in efflux to type IV sera compared with type IIb or NLP. In addition, type IV sera exhibited two-fold higher pre-beta HDL relative concentration (percentage of total apo AI) compared with NLP. Moreover, positive correlations were established between ABCA1-mediated efflux and the serum pre-beta HDL levels or TG concentrations. Thus, the hyperTGemia is associated with a higher fraction of apo AI recovered as pre-beta HDL which appear to be partly responsible for enhanced efflux obtained upon the cAMP stimulation of J774 cells. In conclusion, we demonstrated for the first time that the ABCA1-expressing J774 cell system is responsive to the percent of apo AI present in human serum as pre-beta HDL. Our results suggest that high-plasma TG, accompanied by low HDL may not result in an impaired cholesterol efflux capacity.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/metabolismo , Colesterol/metabolismo , AMP Cíclico/metabolismo , Hipertrigliceridemia/sangre , Macrófagos/metabolismo , Transportadoras de Casetes de Unión a ATP/análisis , Adulto , Anciano , Animales , Transporte Biológico/efectos de los fármacos , Western Blotting , Estudios de Casos y Controles , Movimiento Celular , Células Cultivadas , AMP Cíclico/farmacología , Humanos , Hipertrigliceridemia/diagnóstico , Lipoproteínas HDL/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Probabilidad , ARN Mensajero/análisis , Muestreo , Sensibilidad y Especificidad , Regulación hacia ArribaRESUMEN
Exhaled nitric oxide (NO) concentration, a marker of pulmonary inflammation, has been shown to be elevated in various models of acute lung injury (ALI). This study was undertaken to evaluate the pulmonary NO production in a rat model of postextracorporeal circulation (ECC) ALI. Wistar rats underwent either a partial femorofemoral ECC in normothermia for 3 h (n = 10) or a sham procedure (n = 10). The extracorporeal circuit consisted of a roller pump and a membrane oxygenator. Exhaled NO concentration was monitored with a chemiluminescence analyzer. After sacrifice, lungs were harvested for microscopic studies and to analyze the inducible nitric oxide synthase (iNOS) activity and expression (Western blot). ECC was responsible for an ALI characterized by a decreased arterial blood oxygen saturation (88.9% [51.7-94.2] vs. 93.7% [91.4-98.6] P = 0.005) and pulmonary histological changes (marked alveolar neutrophil infiltration; interstitial edema; intraalveolar hemorrhage). The lung injury score was significantly higher in the ECC group (n = 5; 3.0 [2-4]) in comparison to the sham group (n = 5; 1.0 [0-2]). Exhaled NO concentration remained stable throughout the experiment in all sham rats whereas it significantly increased in the ECC group from baseline (2 ppb [1-5]) until the end of experiment (33.5 ppb [1-47]). Lung iNOS activity and expression were also significantly increased in the ECC group. An increase in exhaled NO, however, did not correlate with the decrease in arterial oxygen pressure. ECC was responsible for an ALI in rats and for an elevated pulmonary NO production. Determination of the relationship between exhaled NO and the severity of the inflammatory process in ALI will require further studies.
Asunto(s)
Circulación Extracorporea , Óxido Nítrico/metabolismo , Enfermedad Aguda , Animales , Western Blotting , Pruebas Respiratorias , Calcio/metabolismo , Espiración , Inflamación , Mediciones Luminiscentes , Pulmón/enzimología , Masculino , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico Sintasa de Tipo II , Oxígeno/metabolismo , Presión , Ratas , Ratas Wistar , Temperatura , Factores de TiempoRESUMEN
OBJECTIVE: This study was undertaken to determine whether an increase in exhaled nitric oxide would reflect the severity of the acute lung injury caused by extracorporeal circulation. DESIGN: Prospective, controlled animal laboratory investigation. SETTING: University laboratory. SUBJECTS: Male, anesthetized, paralyzed, and mechanically ventilated Wistar rats (n = 34). INTERVENTIONS: Twenty-three Wistar rats underwent a partial (100 mL.kg(-1).min(-1)) femoro-femoral extracorporeal circulation in normothermia for 90 mins. Eleven time-matched rats formed the sham group. MEASUREMENTS AND MAIN RESULTS: Exhaled nitric oxide was monitored with a chemiluminescence analyzer. Acute lung injury was assessed by blood gas analysis and lung water content. Lung Evans blue dye content, lung myeloperoxidase, and heme oxygenase activities were determined. Compared with the sham rats, extracorporeal circulation was responsible for acute lung injury characterized by an increased lung water content (82.4 +/- 1.3% vs. 77.9 +/- 1.1%; p<.05), an increased Evans blue dye content (191.8 +/- 15.8 vs. 112.5 +/- 16.8 mg/g tissue wet weight; <.01), and an increased pulmonary heme oxygenase activity (332.9 +/- 107 vs. 113.7 +/- 46.5 pmol. hr(-1).mg of protein(-1); p<.05). Exhaled nitric oxide remained stable throughout the experiment in all sham rats. Among the 23 rats that underwent extracorporeal circulation, eight rats (35%) experienced an increase in exhaled nitric oxide concentration (16.9 +/- 12.7 ppb). There was no significant difference between rats that did or did not experience an increase in exhaled nitric oxide regarding each index of acute lung injury. CONCLUSIONS: An increase in exhaled nitric oxide did not reflect the severity of the acute lung injury caused by extracorporeal circulation. Its significance remains to be determined.