RESUMEN
The enzyme activation-induced deaminase (AID) deaminates deoxycytidine at the immunoglobulin genes, thereby initiating antibody affinity maturation and isotype class switching during immune responses. In contrast, off-target DNA damage caused by AID is oncogenic. Central to balancing immunity and cancer is AID regulation, including the mechanisms determining AID protein levels. We describe a specific functional interaction between AID and the Hsp40 DnaJa1, which provides insight into the function of both proteins. Although both major cytoplasmic type I Hsp40s, DnaJa1 and DnaJa2, are induced upon B-cell activation and interact with AID in vitro, only DnaJa1 overexpression increases AID levels and biological activity in cell lines. Conversely, DnaJa1, but not DnaJa2, depletion reduces AID levels, stability and isotype switching. In vivo, DnaJa1-deficient mice display compromised response to immunization, AID protein and isotype switching levels being reduced by half. Moreover, DnaJa1 farnesylation is required to maintain, and farnesyltransferase inhibition reduces, AID protein levels in B cells. Thus, DnaJa1 is a limiting factor that plays a non-redundant role in the functional stabilization of AID.
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Citidina Desaminasa/metabolismo , Proteínas del Choque Térmico HSP40/metabolismo , Animales , Línea Celular Tumoral , Femenino , Proteínas del Choque Térmico HSP40/genética , Humanos , Masculino , Ratones , Ratones Noqueados , Microscopía ConfocalRESUMEN
Activation-induced deaminase converts deoxycytidine to deoxyuridine at the Ig loci. Complementary pathways, initiated by the uracil-DNA glycosylase (UNG) or the mismatch repair factor MSH2/MSH6, must process the deoxyuridine to initiate class-switch recombination (CSR) and somatic hypermutation. UNG deficiency most severely reduces CSR efficiency and only modestly affects the somatic hypermutation spectrum in vitro. This would predict isotype-switching deficiency but normal affinity maturation in Ung(-/-) mice in vivo, but this has not been tested. Moreover, puzzling differences in the amount of circulating Ig between UNG-deficient humans and mice make it unclear to what extent MSH2/MSH6 can complement for UNG in vivo. We find that Ab affinity maturation is indeed unaffected in Ung(-/-) mice, even allowing IgM responses with higher than normal affinity. Ung(-/-) mice display normal to only moderately reduced basal levels of most circulating Ig subclasses and gut-associated IgA, which are elicited in response to chronically available environmental Ag. In contrast, their ability to produce switched Ig in response to immunization or vesicular stomatitis virus infection is strongly impaired. Our results uncover a specific need for UNG in CSR for timely and efficient acute Ab responses in vivo. Furthermore, Ung(-/-) mice provide a novel model for separating isotype switching and affinity maturation during acute (but not chronic) Ab responses, which could be useful for dissecting their relative contribution to some infections. Interestingly, Ung(-/-) mice present with circulating autoantibodies, suggesting that UNG may impinge on tolerance.
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Afinidad de Anticuerpos/inmunología , Linfocitos B/inmunología , Cambio de Clase de Inmunoglobulina/inmunología , Activación de Linfocitos/inmunología , Uracil-ADN Glicosidasa/inmunología , Animales , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Linfocitos B/metabolismo , Diferenciación Celular/inmunología , Ensayo de Inmunoadsorción Enzimática , Inmunohistoquímica , Inmunoprecipitación , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Uracil-ADN Glicosidasa/deficienciaRESUMEN
Objectives: The incidence of very-early-onset inflammatory bowel disease (VEO-IBD) and early-onset IBD (EO-IBD) is increasing. Here, we report their phenotype and outcomes in a Montreal pediatric cohort. Methods: We analyzed data from patients diagnosed with IBD between January 2014 and December 2018 from the CHU Sainte-Justine. The primary endpoint was to compare the phenotypes of VEO-IBD and EO-IBD. The secondary endpoints involved comparing outcomes and rates of steroid-free clinical remission (SFCR) at 12 (±2) months (m) post-diagnosis and at last follow-up. Results: 28 (14 males) and 67 (34 males) patients were diagnosed with VEO-IBD and EO-IBD, respectively. Crohn's disease (CD) was more prevalent in EO-IBD (64.2% vs. 39.3%), whereas unclassified colitis (IBD-U) was diagnosed in 28.6% of VEO-IBD vs. 10.4% of EO-IBD (p < 0.03). Ulcerative colitis (UC) and IBD-U predominantly presented as pancolitis in both groups (VEO-IBD: 76.5% vs. EO-IBD: 70.8%). Combining all disease subtypes, histological upper GI lesions were found in 57.2% of VEO-IBD vs. 83.6% of EO-IBD (p < 0.009). In each subtype, no differential histological signature (activity, eosinophils, apoptotic bodies, granulomas) was observed between both groups. At 12â m post-diagnosis, 60.8% of VEO-IBD and 62.7% of EO-IBD patients were in SFCR. At a median follow-up of 56â m, SFCR was observed in 85.7% of VEO-IBD vs. 85.0% of EO-IBD patients. Conclusion: The rate of patients in SFCR at 1-year post-diagnosis and at the end of follow-up did not significantly differ between both groups.
RESUMEN
BACKGROUND: Two atherosclerosis-susceptible and -resistant Japanese quail (Coturnix japonica) strains obtained by divergent selection are commonly used as models to study atherosclerosis, but no genetic characterization of their phenotypic differences has been reported so far. Our objective was to examine possible differences in the expression of genes involved in cholesterol metabolism and transport in the liver between these two strains and to evaluate the value of this model to analyze the gene system affecting cholesterol metabolism and transport. METHODS: A factorial study with both strains (atherosclerosis-susceptible versus atherosclerosis-resistant) and two diets (control versus cholesterol) was carried out. The mRNA concentrations of four genes involved in cholesterol biosynthesis (HMGCR, FDFT1, SQLE and DHCR7) and three genes in cholesterol transport (ABCG5, ABCG8 and APOA1) were assayed using real-time quantitative PCR. Plasma lipids were also assayed. RESULTS: Expression of ABCG5 (control diet) and ABCG8 (regardless of dietary treatment) and expression of HMGCR, FDFT1 and SQLE (regardless of dietary treatment) were significantly higher in the atherosclerosis-resistant than in the atherosclerosis-susceptible strain. Plasma triglyceride and LDL levels, and LDL/HDL ratio were significantly higher in the atherosclerosis-susceptible than in the atherosclerosis-resistant strain fed the cholesterol diet. In the atherosclerosis-susceptible strain, ABCG5 expression regressed significantly and positively on plasma LDL level, whereas DHCR7 and SQLE expression regressed significantly and negatively on plasma triglyceride level. CONCLUSIONS: Our results provide support for the hypothesis that the atherosclerosis-resistant strain metabolizes and excretes cholesterol faster than the atherosclerosis-susceptible strain. We have also demonstrated that these quail strains are a useful model to study cholesterol metabolism and transport in relation with atherosclerosis.
Asunto(s)
Aterosclerosis/genética , Colesterol/metabolismo , Coturnix/genética , Coturnix/metabolismo , Resistencia a la Enfermedad/genética , Perfilación de la Expresión Génica , Hígado/metabolismo , Animales , Aterosclerosis/sangre , Transporte Biológico/genética , Colesterol/sangre , Coturnix/clasificación , Dieta , Susceptibilidad a Enfermedades/sangre , Regulación de la Expresión Génica , Lipoproteínas HDL/sangre , Lipoproteínas LDL/sangre , Hígado/patología , Masculino , Ácido Mevalónico/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Regresión , Transducción de Señal/genética , Triglicéridos/sangreRESUMEN
Background and Aim: Patients with inflammatory bowel disease (IBD) are at increased risk for life-threatening complications of Epstein-Barr virus (EBV), including lymphoproliferative diseases. These complications are likely related to inherent immune dysfunction and immunomodulating therapies often used. We aimed to determine the seroprevalence of EBV at diagnosis in our population, its impact on disease at onset, and the risk of active EBV infection. Methods: We included patients newly diagnosed with IBD for whom an EBV serology was performed over a 2-year period. Demographic information and data on disease characteristics were collected retrospectively. Stored serum from the time of diagnosis was retrieved when available for the patients with positive EBV serology, and quantitative polymerase chain reaction testing was performed to assess the pre-treatment viral load of EBV. Results: One hundred twenty patients were included in the study. Fifty-three patients (44.2%) had positive EBV serology at diagnosis. Stratified by age group, the prevalence of seropositive patients was for 0 to <10 years 35%, 10 to <17 years 46%, and ≥17 years 50%. Overall, therapies started within 6 months of diagnosis were similar in both the seropositive and seronegative groups. Within the seropositive group, 66% received systemic corticosteroids, 32.1% infliximab, 5.7% adalimumab, and 5.7% azathioprine. Conclusion: EBV seroprevalence is high in pediatric patients with IBD. EBV seropositivity did not seem to influence the severity of disease at onset or initial choice of therapy.
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SUMMARY: Sarcoidosis is a multisystem disease with various clinical manifestations. It is characterized primarily on a histopathologic basis by the presence of noncaseating granulomata. Laryngeal involvement reportedly occurs in 3-5% of cases, and it is typically localized to the supraglottic region. Patients often present with hoarseness, dysphagia, stridor, or dyspnea. Laryngoscopy typically demonstrates a pale, edematous epiglottis studded with nodules. Tissue biopsy reveals the classic noncaseating granuloma; however, the finding is not diagnostic. Sarcoidosis remains a diagnosis of exclusion to be entertained once other verifiable etiologies for granulomatous disease of the larynx, such as TB, syphilis, fungal infection, berylliosis, or Wegener's granulomatosis, have been ruled out. Systemic corticosteroids remain the mainstay of treatment; however, new steroid-sparing therapies that target the inflammatory response of sarcoidosis are currently being investigated. The case history of a patient with laryngeal sarcoidosis who was managed with the immunosuppressant azathioprine (Imuran) is summarized along with a discussion of other treatment options.
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Azatioprina/uso terapéutico , Citotoxinas/uso terapéutico , Inmunosupresores/uso terapéutico , Enfermedades de la Laringe/tratamiento farmacológico , Sarcoidosis/tratamiento farmacológico , Adulto , Negro o Afroamericano , Azatioprina/administración & dosificación , Citotoxinas/administración & dosificación , Humanos , Inmunosupresores/administración & dosificación , Enfermedades de la Laringe/diagnóstico , Enfermedades de la Laringe/patología , Masculino , Sarcoidosis/diagnóstico , Sarcoidosis/patología , Resultado del TratamientoRESUMEN
OBJECTIVES: The introduction of 48-hour wireless pH testing provides a novel technique of evaluating persons with suspected reflux disease. The wireless capsule can be placed in a sedated individual at the time of esophagogastroduodenoscopy (EGD) or in an unsedated individual at a time after the initial EGD, at the time of esophageal manometry or at the time of transnasal esophagoscopy. The effect that sedation has on the results of 48-hour wireless pH testing has not been evaluated. PURPOSE: To evaluate the day to day variability and the effect of sedation on the results of 48-hour wireless pH testing. METHODOLOGY: The charts of all patients at a tertiary swallowing center undergoing 48-hour wireless pH testing between June 1, 2003 and December 31, 2004 were retrospectively evaluated. Data concerning study indications, route of pH capsule placement, duration of pH recording, and test results were collected. Day to day variability was evaluated, and the results obtained from persons with sedated and unsedated pH capsule placement were compared. RESULTS: Two hundred and six studies were performed. The indications for the examination were gastroesophageal reflux disease (146/206), chronic cough (36/206), and laryngopharyngeal reflux (24/206). Sixty-two percent (128/206) of the studies were performed without sedation and 38% (78/206) with sedation. The overall reproducibility of the daily pH recordings (day 1 vs. day 2) was 77%. Although some trends were observed, there was no significant association between the use of sedation and any of the reflux parameters on pH testing (P > .05). CONCLUSIONS: The day to day reproducibility of 48-hour wireless pH testing was 77%. Intravenous sedation does not appear to have a significant effect on the results of 48-hour wireless pH testing.
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Reflujo Gastroesofágico/diagnóstico , Monitoreo Ambulatorio/métodos , Oligopéptidos , Telemetría/métodos , Cápsulas , Tos/diagnóstico , Endoscopía del Sistema Digestivo , Esofagoscopía , Femenino , Humanos , Concentración de Iones de Hidrógeno , Masculino , Manometría , Enfermedades Faríngeas/diagnóstico , Reproducibilidad de los Resultados , Estudios RetrospectivosRESUMEN
Endotracheal and endobronchial schwannomas are extremely rare tumors of neurogenic origin. These tumors often present late. Common symptoms of hemoptysis and dyspnea result from the size and location of the tumors. Two cases of tracheobronchial schwannoma with proximal airway obstruction are reported. Various diagnostic modalities employed in the evaluation of these tumors are reviewed. Because malignant transformation is exceedingly rare, operative resection of tracheobronchial schwannomas based on the principle of conservation of lung parenchyma is emphasized.
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Obstrucción de las Vías Aéreas/etiología , Neurilemoma/diagnóstico , Neoplasias de la Tráquea/diagnóstico , Adulto , Biopsia , Broncoscopía , Diagnóstico Diferencial , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Neurilemoma/complicaciones , Neurilemoma/cirugía , Tomografía Computarizada por Rayos X , Neoplasias de la Tráquea/complicaciones , Neoplasias de la Tráquea/cirugíaRESUMEN
OBJECTIVE: Experimental study has shown that myocardial ischemic tolerance is reduced during middle-age. We investigated the effect of propofol on ischemic tolerance of middle-aged rat hearts. METHODS: Hearts of young adult (10 weeks old, Y) and middle-aged rats (20 weeks old, M) were assigned to propofol (P-Y, P-M) and control (C-Y, C-M) groups (n=6 each). Hearts were perfused using a Langendorff preparation with Krebs-Henseleit solution (KH) at constant flow rates. We applied propofol (P-Y, P-M) for 10 min at 12 microg/ml before inducing 40 min global ischemia. During ischemia, saline (C-Y, C-M) or propofol (P-Y, P-M) in saline was perfused through the aorta at 60 microl/min. Propofol in KH was perfused at 12 microg/ml for the first 15 min of reperfusion and subsequently reduced to 5 microg/ml in propofol treatment groups. Coronary effluent was assayed for 15-F(2t)-isoprostane after equilibration, during ischemia (T(1)) and at 0.5 (T(2)) and 5 (T(3)) min of reperfusion. After 90 min of reperfusion (T(4)), hearts were harvested to assess tissue antioxidant capacity. RESULTS: In P-Y, we observed an increased latency to ischemic-contracture and a significantly reduced contracture after 35 min ischemia compared to control groups. No ischemic contracture was observed in P-M. There were significantly lower 15-F(2t)-isoprostane levels in P-M and P-Y than in C-M and C-Y at T(1). At T(4), the recovery of left ventricular developed pressure in P-M was greater than in P-Y (P<0.05); both were greater than in C-M and C-Y. CONCLUSION: Propofol enhanced ischemic tolerance of middle-aged hearts, primarily by inhibiting lipid peroxidation.
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Envejecimiento , Antioxidantes/uso terapéutico , Daño por Reperfusión Miocárdica/prevención & control , Propofol/uso terapéutico , Animales , Antioxidantes/metabolismo , Susceptibilidad a Enfermedades , Masculino , Isquemia Miocárdica/metabolismo , Miocardio/metabolismo , Oxidantes/farmacología , Perfusión , Propofol/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Sustancias Reactivas al Ácido Tiobarbitúrico/análisis , terc-Butilhidroperóxido/farmacologíaRESUMEN
BACKGROUND: The role of platelet activating factor (PAF) and nitric oxide in myocardial ischemia-reperfusion (MIR) injury and the interrelationship of the two mediators is poorly understood. The contribution of PAF to apoptosis during MIR has not been studied. OBJECTIVES: To determine the contribution of PAF to ex vivo cardiac dysfunction during the initial 5 h of postischemia reperfusion, to determine the contribution of PAF to inducible nitric oxide synthase (NOS) and endothelial NOS mRNA expression during MIR, and to determine whether PAF contributes to apoptosis during MIR. METHODS: Isolated blood-perfused rabbit hearts underwent 30 min of global ischemia and 5 h reperfusion. Animals were divided into four groups, which received either PAF antagonist TCV-309 or vehicle before ischemia, or were sham operated (heart perfusion only), or were control (no heart perfusion). RESULTS: Administration of the PAF antagonist significantly improved myocardial contractility (614 mmHg/s versus 308 mmHg/s, positive dP/dt, P<0.0001) and coronary vascular flow rate (5.5 mL/min versus 3.9 mL/min, P<0.01) during reperfusion compared with untreated animals (values at 5 h reperfusion). Treatment with PAF antagonist significantly increased mRNA expression of endothelial NOS (2.8 versus 1.3 ratio, P<0.05) compared with the untreated group. PAF antagonist reduced procaspase-3 cleavage (66 versus 108 ratio, P<0.05) and DNA fragmentation (8.2 versus 11.0 positive cells per field) compared with untreated animals. CONCLUSIONS: PAF antagonism with TCV-309 protected against myocardial contractile depression and coronary vasoconstriction during the initial 5 h reperfusion. PAF may play a role in the regulation of endothelial NOS mRNA expression and contribute to apoptosis during ischemia-reperfusion in the heart.
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Apoptosis , Contracción Miocárdica , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/fisiopatología , Óxido Nítrico Sintasa/metabolismo , Factor de Activación Plaquetaria/metabolismo , Tetrahidroisoquinolinas , Animales , Apoptosis/efectos de los fármacos , Western Blotting , Circulación Coronaria/efectos de los fármacos , Fragmentación del ADN , Expresión Génica/efectos de los fármacos , Técnicas In Vitro , Isoquinolinas/farmacología , Contracción Miocárdica/efectos de los fármacos , Daño por Reperfusión Miocárdica/enzimología , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa de Tipo II , Óxido Nítrico Sintasa de Tipo III , Factor de Activación Plaquetaria/antagonistas & inhibidores , Inhibidores de Agregación Plaquetaria/farmacología , Compuestos de Piridinio/farmacología , ARN Mensajero/metabolismo , Conejos , Factores de TiempoRESUMEN
Reactive oxygen species induce formation of 15-F(2t)-isoprostane (15-F(2t)-IsoP), a specific marker of in vivo lipid peroxidation, which is increased after myocardial ischemia and during the subsequent reperfusion. 15-F(2t)-IsoP possesses potent bioactivity under pathophysiological conditions. However, it remains unknown whether 15-F(2t)-IsoP, by itself, can influence myocardial ischemia-reperfusion injury (IRI). Adult rat hearts were perfused by the Langendorff technique with Krebs-Henseleit (KH) solution at a constant flow rate of 10 ml/min. 15-F(2t)-IsoP (100 nM), SQ-29548 (1 microM, SQ), a thromboxane receptor antagonist that can abolish the vasoconstrictor effect of 15-F(2t)-IsoP, 15-F(2t)-IsoP + SQ in KH, or KH alone (vehicle control) was applied for 10 min before induction of 40 min of global ischemia followed by 60 min of reperfusion. During ischemia, saline (control), 15-F(2t)-IsoP, 15-F(2t)-IsoP + SQ, or SQ in saline was perfused through the aorta at 60 microl/min. 15-F(2t)-IsoP, 15-F(2t)-IsoP + SQ, or SQ in KH was infused during the first 15 min of reperfusion. Coronary effluent endothelin-1 concentrations were significantly higher in the group treated with 15-F(2t)-IsoP than in the control group during ischemia and also in the later phase of reperfusion (P < 0.05). Infusion of 15-F(2t)-IsoP increased release of cardiac-specific creatine kinase, reduced cardiac contractility during reperfusion, and increased myocardial infarct size relative to the control group. SQ abolished the deleterious effects of 15-F(2t)-IsoP. 15-F(2t)-IsoP exacerbates myocardial IRI and may, therefore, act as a mediator of IRI. 15-F(2t)-IsoP-induced endothelin-1 production during cardiac reperfusion may represent a mechanism underlying the deleterious actions of 15-F(2t)-IsoP.
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Dinoprost/análogos & derivados , Corazón/efectos de los fármacos , Daño por Reperfusión Miocárdica/metabolismo , Miocardio/metabolismo , Vasoconstrictores/farmacología , Animales , Presión Sanguínea/fisiología , Creatina Quinasa/metabolismo , Forma MB de la Creatina-Quinasa , Dinoprost/farmacología , Endotelina-1/metabolismo , Corazón/fisiología , Técnicas In Vitro , Isoenzimas/metabolismo , Masculino , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Infarto del Miocardio/fisiopatología , Daño por Reperfusión Miocárdica/patología , Daño por Reperfusión Miocárdica/fisiopatología , Miocardio/patología , Ratas , Ratas Sprague-DawleyRESUMEN
Several lines of evidence suggest that antioxidant processes and (or) endogenous antioxidants inhibit proatherogenic events in the blood vessel wall. Heme oxygenase (HO), which catabolizes heme to biliverdin, carbon monoxide, and catalytic iron, has been shown to have such antioxidative properties. The HO-1 isoform of heme oxygenase is ubiquitous and can be increased several fold by stimuli that induce cellular oxidative stress. Products of the HO reaction have important effects: carbon monoxide is a potent vasodilator, which is thought to play a role in modulation of vascular tone; biliverdin and its by-product bilirubin are potent antioxidants. Although HO induction results in an increase in catalytic free iron release, the enhancement of intracellular ferritin protein through HO-1 has been reported to decrease the cytotoxic effects of iron. Oxidized LDL has been shown to increase HO-1 expression in endothelial and smooth muscle cell cultures, and during atherogenesis. Further evidence of HO-1 expression associated with atherogenesis has been demonstrated in human, murine and rabbit atherosclerotic lesions. Moreover, genetic models of HO deficiency suggest that the actions of HO-1 are important in modulating the severity of atherosclerosis. Recent experiments in gene therapy using the HO gene suggest that interventions aimed at HO in the vessel wall could provide a novel therapeutic approach for the treatment or prevention of atherosclerotic disease.
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Arteriosclerosis/metabolismo , Vasos Sanguíneos/enzimología , Endotelio Vascular/enzimología , Regulación Enzimológica de la Expresión Génica , Hemo Oxigenasa (Desciclizante)/genética , Hemo Oxigenasa (Desciclizante)/metabolismo , Animales , Antioxidantes/metabolismo , Arteriosclerosis/fisiopatología , Arteriosclerosis/terapia , Bilirrubina/metabolismo , Biliverdina/metabolismo , Vasos Sanguíneos/patología , Monóxido de Carbono/metabolismo , Endotelio Vascular/patología , Ferritinas/metabolismo , Terapia Genética , Hemo/metabolismo , Humanos , Isoenzimas/genética , Isoenzimas/metabolismo , Lipoproteínas LDL/metabolismo , Estrés OxidativoRESUMEN
Previous studies have shown that reactive oxygen species mediated lipid peroxidation in patients undergoing cardiac surgery occurs primarily during cardiopulmonary bypass. We examined whether application of a high concentration of propofol during ischemia could effectively enhance postischemic myocardial functional recovery in the setting of global ischemia and reperfusion in an isolated heart preparation. Hearts were subjected to 40 min of global ischemia followed by 90 min of reperfusion. During ischemia, propofol (12 microg/mL in saline) was perfused through the aorta at 60 microL/min. We found that application of high-concentration propofol during ischemia combined with low-concentration propofol (1.2 microg/mL) administered before ischemia and during reperfusion significantly improved postischemic myocardial functional recovery without depressing cardiac mechanics before ischemia, as is seen when high-concentration propofol was applied prior to ischemia and during reperfusion. The functional enhancement is associated with increased heart tissue antioxidant capacity and reduced lipid peroxidation. We conclude that high-concentration propofol application during ischemia could be a potential therapeutic and anesthetic strategy for patients with preexisting myocardial dysfunction.
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Antioxidantes/metabolismo , Corazón/efectos de los fármacos , Isquemia Miocárdica/tratamiento farmacológico , Miocardio/metabolismo , Propofol/administración & dosificación , Animales , Corazón/fisiología , Técnicas In Vitro , Masculino , Isquemia Miocárdica/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
UNLABELLED: The effects of anesthesia on ischemia-reperfusion injury are of considerable scientific and clinical interest. We examined the effects of propofol (known to possess antioxidant activity) and halothane (devoid of antioxidant activity in vitro) on tissue and red blood cell (RBC) antioxidant capacity. Adult male Wistar rats were anesthetized with halothane 0.5%-1.0% (n = 7), propofol 500 microg x kg(-1) x min(-1) with halothane 0.25%-0.5% (small-dose propofol; n = 9), or propofol 2000 microg x kg(-1) x min(-1) (large-dose propofol; n = 8) for 45 min. Blood and tissue samples of liver, kidney, heart, and lung were then harvested for in vitro exposure to a peroxidizing agent. Red cell malondialdehyde and tissue thiobarbituric acid reactive substances were determined spectrophotometrically. Antioxidant capacities of blood and tissues in the Large-Dose Propofol group, and of blood and all tissues except lung in the Small-Dose Propofol group, were increased significantly compared with halothane (P < 0.003). The increases in tissue antioxidant capacities varied in their magnitude: RBC > liver > kidney > heart > lung. There was a high correlation between changes in RBC susceptibility to oxidative damage and corresponding changes in tissues. These findings demonstrate that large-dose propofol significantly enhances tissue antioxidant capacity, and RBC antioxidant capacity can serve as a functional measure of tissue activity, in vivo. IMPLICATIONS: We designed this study to investigate the antioxidant effects of propofol in various tissues in a rat model. Pretreatment of animals with propofol led to a reduction in the susceptibility to an in vitro oxidative stress of five different tissues investigated, demonstrating the drug's ability to limit oxidative injury. This may have future application in limiting organ dysfunction after periods of tissue ischemia (which results in oxidative damage).
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Anestésicos Intravenosos/farmacología , Antioxidantes/metabolismo , Eritrocitos/metabolismo , Propofol/farmacología , Análisis de Varianza , Anestésicos por Inhalación , Anestésicos Intravenosos/administración & dosificación , Animales , Relación Dosis-Respuesta a Droga , Halotano/farmacología , Riñón/metabolismo , Hígado/metabolismo , Pulmón/metabolismo , Masculino , Malondialdehído/metabolismo , Miocardio/metabolismo , Propofol/administración & dosificación , Ratas , Ratas Wistar , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , terc-Butilhidroperóxido/farmacologíaRESUMEN
We have investigated heme oxygenase (HO) and antioxidant status in the novel isolation and characterization of aortic endothelial cells (AECs) from a random bred wild-type strain (WILD) and selectively bred atherosclerosis-susceptible (SUS) and -resistant (RES) strains of Japanese quail. Cultured AECs expressed acetylated LDL, and were probed with endothelial and smooth muscle cell specific antibodies to confirm purity of culture. Subconfluent monolayers of RES AECs had higher HO activity than SUS AECs. At confluence, HO activity levels were similar among strains. However, RES AECs had higher HO-1 protein than WILD and SUS cells. Although ferritin protein levels were similar among the three strains, catalytic iron was higher in SUS AECs than WILD and RES cells. Glutathione levels were highest in SUS cells, intermediate in WILD, and lowest in RES, while glutathione reductase was higher in WILD and RES AECs than SUS AECs. We suggest that differences in atherosclerosis susceptibility between RES and SUS may be due, at least in part, to differences in endothelial HO and antioxidant components.
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Antioxidantes/metabolismo , Arteriosclerosis/metabolismo , Endotelio Vascular/metabolismo , Hemo Oxigenasa (Desciclizante)/metabolismo , Animales , Arteriosclerosis/enzimología , Western Blotting , Células Cultivadas , Coturnix , Susceptibilidad a Enfermedades , Electroforesis en Gel de Poliacrilamida , Endotelio Vascular/citología , Endotelio Vascular/enzimología , Ferritinas/metabolismo , Glutatión/metabolismo , Glutatión Peroxidasa/metabolismo , Glutatión Reductasa/metabolismo , Inmunohistoquímica , Superóxido Dismutasa/metabolismoRESUMEN
Recent studies on cultured aortic endothelial cells (AECs) from atherosclerosis-susceptible (SUS) and -resistant (RES) strains of Japanese quail suggest that differences in atherosclerosis susceptibility between RES and SUS may be due to differences in endothelial heme oxygenase (HO) and antioxidant components. We have now investigated the effects of oxidant-induced injury on HO and glutathione (GSH) in AECs from SUS and RES quail. We report that cultured AECs from SUS and RES birds differ in their response to oxidative stress. AECs from the SUS strain cells are more susceptible than those from the RES strain to oxidative stress induced by tert-butylhydroperoxide, as judged by lower HO activity, HO-1 expression, ferritin and GSH levels. Aortic endothelial cells from SUS birds also showed higher levels of catalytic iron, TBARS production and LDH release compared with RES cells, indicating that SUS AECs are more susceptible to oxidative stress than cells from the resistant strain. Furthermore, independently of genetic status, AECs from old birds have higher TBARS and lower levels of HSP70 induction than AECs from younger birds, suggesting that aging is associated with a decreased ability of AECs to respond to oxidative stress, and this may be relevant to the permissive effect of aging on the process of atherogenesis. Our results indicate that genetic factors and endogenous antioxidant systems in the blood vessel wall may be important in determining the susceptibility of vascular cells to oxidative stress and atherosclerotic plaque formation.
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Aorta/citología , Arteriosclerosis/patología , Endotelio Vascular/citología , Glutatión/metabolismo , Lesiones Cardíacas/patología , Hemo Oxigenasa (Desciclizante)/metabolismo , Oxidantes/farmacología , Animales , Antioxidantes/metabolismo , Aorta/patología , Bleomicina/farmacología , Western Blotting , Catálisis , Línea Celular , Células Cultivadas , Coturnix , Endotelio Vascular/metabolismo , Ensayo de Inmunoadsorción Enzimática , Ferritinas/biosíntesis , Predisposición Genética a la Enfermedad , Proteínas HSP70 de Choque Térmico/metabolismo , Hemo-Oxigenasa 1 , Inmunohistoquímica , Hierro/metabolismo , L-Lactato Deshidrogenasa , Peroxidación de Lípido , Estrés Oxidativo , Sustancias Reactivas al Ácido Tiobarbitúrico , Factores de TiempoRESUMEN
We examined the effects of propofol (2,6-diisopropylphenol) on functional recovery and 15-F2t-isoprostane generation during ischemia-reperfusion in Langendorff-perfused rat hearts. Before the induction of 40 min of global ischemia, hearts were perfused (10 min) with propofol at 5 (lo-P) or 12 microg/mL (hi-P) in saline or with saline only (control). During ischemia, saline, lo-P, or hi-P was perfused through the aorta at 60 microL/min. During the first 15 min of reperfusion, propofol (5 or 12 microg/mL) was continued, followed by perfusion with 5 microg/mL propofol for 75 min in both propofol-treated groups. After 90 min of reperfusion (Rep-90), heart tissues were harvested for assessment of antioxidant status. In hi-P, we observed increased latency to and greater reduction of ischemic contracture relative to the lo-P or control groups. 15-F2t-Isoprostane concentrations increased during ischemia and were significantly lower in hi-P and lo-P than in control (P < 0.01). At Rep-90, myocardial functional recovery was greater in both propofol-treated groups relative to control, and it correlated positively with tissue antioxidant capacity preservation. Tissue antioxidant capacity was better preserved in hi-P than in lo-P treatment (P < 0.05). We conclude that oxidant injury occurs during ischemia and reperfusion, and propofol provides dose-dependent protection primarily by enhancing tissue antioxidant capacity and reducing lipid peroxidation.
Asunto(s)
Anestésicos Intravenosos/farmacología , Dinoprost/análogos & derivados , F2-Isoprostanos/biosíntesis , Isquemia Miocárdica/prevención & control , Daño por Reperfusión Miocárdica/prevención & control , Propofol/farmacología , Anestésicos Intravenosos/administración & dosificación , Animales , Antioxidantes/metabolismo , Relación Dosis-Respuesta a Droga , Corazón/efectos de los fármacos , Corazón/fisiopatología , Técnicas In Vitro , Peroxidación de Lípido , Masculino , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/fisiopatología , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/fisiopatología , Propofol/administración & dosificación , Ratas , Ratas Sprague-Dawley , Función Ventricular Izquierda/efectos de los fármacosRESUMEN
Antioxidant component alterations in the aorta during atherogenesis were examined in atherosclerosis-susceptible (SUS) Japanese quail fed a cholesterol-supplemented (0.5% w/w) diet. Birds fed a non-supplemented diet provided information on the effects of aging on endogenous antioxidants. One hundred adult SUS males were used. Birds were sacrificed after 0, 4, 8 and 12 weeks on the diets and were examined for plaque development and corresponding antioxidant component alterations in aorta and myocardium. With aging, superoxide dismutase (SOD) activity was increased in both tissues, whereas aortic glutathione peroxidase (GPx) activity and myocardial glutathione reductase (GRd) activity decreased. Myocardial ascorbate levels increased with aging, with a reciprocal decrease in myocardial tocopherol levels. Following 4 weeks of cholesterol supplementation, aortic GRd decreased, SOD activity increased, but activities of GPx and catalase were unchanged. This same qualitative pattern of antioxidant enzyme changes was also found in myocardium. Thus, although aortic antioxidant enzyme changes produced by cholesterol feeding and aging showed some similarities, the early phase of atherogenesis does not simply reflect accelerated aging. In the late stages of atherogenesis, SOD activity returned to baseline, but other antioxidant enzymes remained unaltered from levels characterizing the early phase of lesion development. There was no detectable functional coupling between changes in GPx and GRd, nor between SOD (which produces hydrogen peroxide) and GPx or catalase (which utilize hydrogen peroxide as substrate). Previously reported alterations in erythrocyte antioxidant enzyme components during atherogenesis in quail were not predictive of changes in the corresponding enzymes in the aorta and myocardium.