RESUMEN
BACKGROUND: X-linked hypohidrotic ectodermal dysplasia (XLHED) is caused by pathogenic variants of the gene EDA disrupting the prenatal development of ectodermal derivatives. Cardinal symptoms are hypotrichosis, lack of teeth, and hypo- or anhidrosis, but the disease may also evoke other clinical problems. This study aimed at investigating the clinical course of XLHED in early childhood as the basis for an evaluation of the efficacy of potential treatments. METHODS: 25 children (19 boys and 6 girls between 11 and 35 months of age) with genetically confirmed XLHED were enrolled in a long-term natural history study. Clinical data were collected both retrospectively using parent questionnaires and medical records (pregnancy, birth, infancy) and prospectively until the age of 60 months. General development, dentition, sweating ability, ocular, respiratory, and skin involvement were assessed by standardized clinical examination and yearly quantitative surveys. RESULTS: All male subjects suffered from persistent anhidrosis and heat intolerance, although a few sweat ducts were detected in some patients. Sweating ability of girls with XLHED ranged from strongly reduced to almost normal. In the male subjects, 1-12 deciduous teeth erupted and 0-8 tooth germs of the permanent dentition became detectable. Tooth numbers were higher but variable in the female group. Most affected boys had no more than three if any Meibomian glands per eyelid, most girls had fewer than 10. Many male subjects developed additional, sometimes severe health issues, such as obstructive airway conditions, chronic eczema, or dry eye disease. Adverse events included various XLHED-related infections, unexplained fever, allergic reactions, and retardation of psychomotor development. CONCLUSIONS: This first comprehensive study of the course of XLHED confirmed the early involvement of multiple organs, pointing to the need of early therapeutic intervention.
Asunto(s)
Displasia Ectodermal Anhidrótica Tipo 1/genética , Antropometría , Preescolar , Displasia Ectodermal Anhidrótica Tipo 1/metabolismo , Displasia Ectodermal Anhidrótica Tipo 1/patología , Ectodisplasinas/genética , Ectodisplasinas/metabolismo , Femenino , Genotipo , Humanos , Lactante , Masculino , Estudios Retrospectivos , Encuestas y CuestionariosRESUMEN
Here we investigated the role of NFAT-interacting protein (NIP)-45, an Interleukin (IL)-4 inducing Transcription Factor, and its impact on the differentiation of Group 2 Innate -Lymphoid -Cells (ILC2s) in the pathogenesis of asthma. NIP45, a transcription factor regulating NFATc1 activity, mRNA was found to be induced in the Peripheral Blood mononuclear cells (PMBCs) of asthmatic pre-school children with allergies and in the peripheral blood CD4+ T cells from adult asthmatic patients. In PBMCs of asthmatic and control children, NIP45 mRNA directly correlated with NFATc1 but not with T-bet. Targeted deletion of NIP45 in mice resulted in a protective phenotype in experimental asthma with reduced airway mucus production, airway hyperresponsiveness and eosinophils. This phenotype was reversed by intranasal delivery of recombinant r-IL-33. Consistently, ILC2s and not GATA3+ CD4+ T-cells were decreased in the lungs of asthmatic NIP45-/- mice. Reduced cell number spleen ILC2s could be differentiated from NIP45-/- as compared to wild-type mice after in vivo injection of a microcircle-DNA vector expressing IL-25 and decreased cytokines and ILC2 markers in ILC2 differentiated from the bone marrow of NIP45-/- mice. NIP45 thus emerges as a new therapeutic target for the resolution of the airway pathology, down-regulation of ILC2s and mucus production in asthma.
Asunto(s)
Asma/genética , Proteínas Portadoras/genética , Inmunidad Innata/genética , Factores de Transcripción NFATC/genética , Animales , Asma/metabolismo , Asma/patología , Niño , Preescolar , Modelos Animales de Enfermedad , Femenino , Humanos , Interleucina-33/genética , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Linfocitos/inmunología , Linfocitos/metabolismo , Masculino , Ratones , Ratones Noqueados , Moco/inmunología , Moco/metabolismo , Hipersensibilidad Respiratoria/genética , Hipersensibilidad Respiratoria/inmunología , Proteínas de Dominio T Box/genéticaRESUMEN
Cystic fibrosis (CF) is characterised by chronic airway infection with bacteria and fungi. Infections caused by Scedosporium/Lomentospora species can occur and are difficult to treat. Moulds belonging to the genus Scedosporium/Lomentospora are detected most frequently in respiratory samples of patients with CF, next to Aspergillus spp. Our aim was to define pulmonary fungal infections due to Scedosporium/Lomentospora in CF and to study the antimycotic treatment. In this multicentre study (12 centres; duration January 2008 to December 2014) 31 patients with a lung infection caused by moulds of the genus Scedosporium/Lomentospora were included. 36 courses of antifungal treatment were documented. Scedosporium apiospermum sensu stricto accounted for 48.4% of cases. In 20/31 patients a therapeutic response under antimycotics (median duration 3.9â¯months) was achieved. Triple and double therapy was significantly more effective compared to monotherapy regarding FEV1, radiology, and symptoms. This data suggests that combined treatment is superior to monotherapy in patients with CF.
Asunto(s)
Antifúngicos , Fibrosis Quística , Quimioterapia Combinada/métodos , Infecciones Fúngicas Invasoras , Enfermedades Pulmonares Fúngicas , Scedosporium , Adulto , Antifúngicos/administración & dosificación , Antifúngicos/clasificación , Fibrosis Quística/complicaciones , Fibrosis Quística/microbiología , Fibrosis Quística/terapia , Monitoreo de Drogas/métodos , Monitoreo de Drogas/estadística & datos numéricos , Femenino , Alemania , Humanos , Infecciones Fúngicas Invasoras/diagnóstico , Infecciones Fúngicas Invasoras/tratamiento farmacológico , Pulmón/diagnóstico por imagen , Pulmón/fisiopatología , Enfermedades Pulmonares Fúngicas/diagnóstico , Enfermedades Pulmonares Fúngicas/tratamiento farmacológico , Enfermedades Pulmonares Fúngicas/microbiología , Masculino , Evaluación de Procesos y Resultados en Atención de Salud , Pruebas de Función Respiratoria/métodos , Scedosporium/efectos de los fármacos , Scedosporium/aislamiento & purificación , Tomografía Computarizada por Rayos X/métodosRESUMEN
BACKGROUND: Aspergillus fumigatus is frequently encountered in sputum samples of patients with cystic fibrosis (CF), which traditionally has been interpreted as saprophytic airway colonization. However, this mere bystander role has been challenged by recent data. There is now evidence that Aspergillus fumigatus accelerates the decline of pulmonary function. (1â3)-ß-D-glucan (BDG) and galactomannan (GM) are highly sensitive fungal biomarkers that are used to diagnose invasive fungal disease. However, their diagnostic value in CF patients is largely unknown. METHODS: We conducted a retrospective cohort study on 104 CF patients to determine whether serum BDG and GM levels correlate with parameters such as Aspergillus-positive sputum cultures and lung function. RESULTS: Aspergillus fumigatus was persistently detected in 22 of the 104 CF patients (21%). Mean serum BDG and GM levels in the Aspergillus-positive patients were significantly higher than in those without persistent Aspergillus detection (89 versus 40 pg/ml [p = 0.022] and 0.30 versus 0.15 ODI [p = 0.013], respectively). 27 and 7 patients had elevated BDG (≥ 60 pg/ml) or GM levels (> 0.5 ODI), respectivly. BDG and GM levels showed a significant correlation (p = 0.004). Patients with increased serum concentrations of BDG were more frequently Aspergillus-positive (40.7 versus 14.3%, p = 0.004) and had a significantly lower forced expiratory volume in one second (FEV1) than patients with a normal BDG (61.6 versus 77.1%, p = 0.007). In the multivariate analysis, BDG but not GM or the growth of A. fumigatus, proved to be an independent predictor for the FEV1. CONCLUSIONS: CF patients with persistent Aspergillus detection have elevated BDG and GM levels which ranged between healthy and invasively infected patients. Serum BDG may be superior to GM and fungal culture in predicting an impaired lung function in CF patients.
Asunto(s)
Aspergillus fumigatus , Fibrosis Quística/fisiopatología , Mananos/sangre , Aspergilosis Pulmonar/sangre , beta-Glucanos/sangre , Adolescente , Adulto , Estudios de Casos y Controles , Niño , Preescolar , Estudios de Cohortes , Técnicas de Cultivo , Fibrosis Quística/complicaciones , Femenino , Volumen Espiratorio Forzado , Galactosa/análogos & derivados , Humanos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Proteoglicanos , Aspergilosis Pulmonar/complicaciones , Aspergilosis Pulmonar/diagnóstico , Estudios Retrospectivos , Esputo/microbiología , Adulto JovenRESUMEN
BACKGROUND: In individuals with cystic fibrosis (CF), colonization with methicillin-resistant Staphylococcus aureus (MRSA) was reported to be associated with a deterioration of pulmonary disease as reflected by an accelerated decline in lung function. Thus, an early eradication of MRSA could be beneficial in these patients. Here, we report on an intensified MRSA eradication protocol. METHODS: Since 2012 a protocol for the eradication of newly acquired MRSA has been used in our CF Clinic, combining oral rifampicin and fusidic acid, inhaled vancomycin, nasal mupirocin, local antiseptic treatment and hygienic directives all of which are applied for only 7 days during an inpatient hospital stay. RESULTS: Since 2012 seven patients (3 male, 4 female; age range 4 to 30 years) newly acquired MRSA. In 6 of the 7 patients (86%) successful eradication of MRSA was achieved upon first treatment using the protocol described above. In one patient a second course of treatment was performed which, however, also failed to eliminate the colonizing MRSA. CONCLUSIONS: Our protocol led to an eradication rate of 86%. The impact of each individual component of the protocol remains to be determined.
Asunto(s)
Antibacterianos/uso terapéutico , Fibrosis Quística/complicaciones , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas/tratamiento farmacológico , Administración por Inhalación , Administración Intranasal , Administración Oral , Adolescente , Adulto , Antibacterianos/administración & dosificación , Antiinfecciosos Locales/uso terapéutico , Baños , Niño , Preescolar , Fibrosis Quística/microbiología , Quimioterapia Combinada/métodos , Femenino , Ácido Fusídico/uso terapéutico , Desinfección de las Manos , Humanos , Masculino , Mupirocina/uso terapéutico , Estudios Retrospectivos , Rifampin/uso terapéutico , Infecciones Estafilocócicas/microbiología , Resultado del Tratamiento , Vancomicina/uso terapéutico , Adulto JovenRESUMEN
Aspergillus fumigatus frequently colonizes the airways of patients with cystic fibrosis (CF) and may cause various severe infections, such as bronchitis. Serological data, sputum dependent markers and longitudinal data of treated cases of Aspergillus bronchitis were evaluated for further description of this infection. This study, which comprises three substudies, aimed to analyze epidemiological data of Aspergillus in CF and the entity of Aspergillus bronchitis. In a first step, data of the German Cystic Fibrosis Registry were used to evaluate the frequency of Aspergillus colonization in patients with CF (n = 2599). Then a retrospective analysis of 10 cases of Aspergillus bronchitis was performed to evaluate longitudinal data for lung function and clinical presentation parameters: sputum production, cough and physical capacity. Finally, a prospective cohort study (n = 22) was conducted to investigate serological markers for Aspergillus bronchitis: total serum IgE, specific serum IgE, specific serum IgG, as well as sputum galactomannan, real-time PCR detection of Aspergillus DNA in sputum and fungal cultures. Analysis of the German CF registry revealed an Aspergillus colonization rate of 32.5% among the 2599 patients. A retrospective data analysis of 10 treated cases revealed the clinical course of Aspergillus bronchitis, including repeated positive sputum culture findings for A. fumigatus, no antibiotic treatment response, total serum IgE levels <200 kU/l, no observation of new pulmonary infiltrates and appropriate antifungal treatment response. Antifungal treatment durations of 4 ± 1.6 (2-6) weeks significantly reduced cough (P = 0.0067), sputum production (P < 0.0001) and lung function measures (P = 0.0358) but not physical capacity (P = 0.0794). From this retrospective study, a prevalence of 1.6% was calculated. In addition, two cases of Aspergillus bronchitis were identified in the prospective cohort study according to immunological, molecular and microbiological parameters. A prevalence of 9% was assessed. Aspergillus bronchitis appears to occur in a minority of colonized CF patients. Antifungal treatment may reduce respiratory symptoms and restore lung function.
Asunto(s)
Aspergilosis/epidemiología , Aspergilosis/patología , Aspergillus fumigatus/aislamiento & purificación , Bronquitis/epidemiología , Bronquitis/patología , Fibrosis Quística/complicaciones , Adolescente , Adulto , Antifúngicos/uso terapéutico , Antígenos Fúngicos/análisis , Aspergilosis/diagnóstico , Aspergilosis/tratamiento farmacológico , Biomarcadores/sangre , Bronquitis/diagnóstico , Bronquitis/tratamiento farmacológico , Niño , ADN de Hongos/análisis , Femenino , Galactosa/análogos & derivados , Alemania/epidemiología , Humanos , Estudios Longitudinales , Masculino , Mananos/análisis , Prevalencia , Estudios Prospectivos , Reacción en Cadena en Tiempo Real de la Polimerasa , Pruebas de Función Respiratoria , Estudios Retrospectivos , Suero/química , Esputo/microbiología , Adulto JovenRESUMEN
This corrects the article DOI: 10.1038/srep43426.
RESUMEN
Here we investigated the relationship between local bacterial colonization and anti-bacterial immune responses in pre-school asthmatic and control children within the EU-wide study PreDicta. In this cohort of pre-school asthmatic children, nasopharyngeal colonization with Gram-negative bacteria such as Haemophilus influenzae and Moraxella catarrhalis was found to be associated with the highest interferon beta (IFNß) and IL-33 levels in the nasal pharyngeal fluids (NPF). IL33R-ST2 was found induced in the blood of asthmatic children with additional Gram + bacteria in the nasopharynx (Gr+/-). Furthermore, asthmatic children had more episodes of infection that required antibiotic therapy than the control group. Treatment with antibiotics associated with reduced ST2 in blood cells of both asthmatic and control children and reduced IL-33 levels in the airways of asthmatic children. In the absence of Staphylococcus (S.) aureus in NPF, antibiotic therapy associated with decreased IL-33 levels in the NPF and lower ST2 values in the blood of control children but not of asthmatic children. These data suggest that, in asthmatic children, Gram- bacteria, which persist after antibiotic therapy, contributes to IL-33 locally and associated with Gr + bacteria colonization in the airways, inhibited IFN-ß and in the absence of Staphylococcus (S.) aureus, induced ST2 bearing cells in their blood.
Asunto(s)
Asma/inmunología , Infecciones por Haemophilus/inmunología , Proteína 1 Similar al Receptor de Interleucina-1/inmunología , Interleucina-33/inmunología , Infecciones por Moraxellaceae/inmunología , Infecciones Estafilocócicas/inmunología , Antibacterianos/uso terapéutico , Asma/tratamiento farmacológico , Asma/genética , Asma/microbiología , Broncodilatadores/uso terapéutico , Estudios de Casos y Controles , Niño , Preescolar , Femenino , Fluticasona/uso terapéutico , Regulación de la Expresión Génica , Infecciones por Haemophilus/tratamiento farmacológico , Infecciones por Haemophilus/genética , Infecciones por Haemophilus/microbiología , Haemophilus influenzae/efectos de los fármacos , Haemophilus influenzae/crecimiento & desarrollo , Haemophilus influenzae/inmunología , Humanos , Interferón beta/genética , Interferón beta/inmunología , Proteína 1 Similar al Receptor de Interleucina-1/genética , Interleucina-33/genética , Masculino , Moraxella catarrhalis/efectos de los fármacos , Moraxella catarrhalis/crecimiento & desarrollo , Moraxella catarrhalis/inmunología , Infecciones por Moraxellaceae/tratamiento farmacológico , Infecciones por Moraxellaceae/genética , Infecciones por Moraxellaceae/microbiología , Nasofaringe/efectos de los fármacos , Nasofaringe/crecimiento & desarrollo , Nasofaringe/inmunología , Pruebas de Función Respiratoria , Xinafoato de Salmeterol/uso terapéutico , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/genética , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus aureus/inmunologíaAsunto(s)
Interferones , Rhinovirus , Antivirales , Asma , Genotipo , Humanos , Infecciones por PicornaviridaeAsunto(s)
Asma/etiología , Asma/metabolismo , Interacciones Huésped-Patógeno/inmunología , Infecciones por Picornaviridae/complicaciones , Infecciones por Picornaviridae/inmunología , Rhinovirus/inmunología , Factor de Crecimiento Transformador beta/metabolismo , Animales , Asma/diagnóstico , Asma/terapia , Modelos Animales de Enfermedad , Humanos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/metabolismo , Ratones , Infecciones por Picornaviridae/virología , Carga ViralRESUMEN
We analysed the influence of rhinovirus (RV) in nasopharyngeal fluid (NPF) on type I and III interferon (IFN) responses (e.g. IFN-α and IFN -: λ) and their signal transduction, at baseline and during disease exacerbation, in cohorts of pre-school children with and without asthma.At the time of recruitment into the Europe-wide study PreDicta, and during symptoms, NPF was collected and the local RV colonisation was analysed. Peripheral blood mononuclear cells (PBMCs) were challenged in vitro with RV or not. RNA was analysed by quantitative real-time PCR and gene arrays. Serum was analysed with ELISA for IFNs and C-reactive protein.We found that PBMCs from asthmatic children infected in vitro with the RV1b serotype upregulated MYD88, IRF1, STAT1 and STAT2 mRNA, whereas MYD88, IRF1, STAT1 and IRF9 were predominantly induced in control children. Moreover, during symptomatic visits because of disease exacerbation associated with RV detection in NPF, IFN-α production was found increased, while IFN-λ secretion was already induced by RV in asthmatic children at baseline.During asthma exacerbations associated with RV, asthmatic children can induce IFN-α secretion, indicating a hyperactive immune response to repeated respiratory virus infection.
Asunto(s)
Asma/inmunología , Proteína C-Reactiva/análisis , Interferones/sangre , Leucocitos Mononucleares/virología , Infecciones por Picornaviridae/inmunología , Asma/virología , Células Cultivadas , Preescolar , Progresión de la Enfermedad , Europa (Continente) , Femenino , Humanos , Factor 1 Regulador del Interferón/genética , Interferones/inmunología , Masculino , Factor 88 de Diferenciación Mieloide/genética , Nasofaringe/virología , Estudios Prospectivos , ARN Mensajero/análisis , Rhinovirus , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT2/genética , Transducción de SeñalRESUMEN
BACKGROUND/AIMS: In cystic fibrosis (CF), chronic microbial lung infections are difficult to treat and cause morbidity and increased mortality. METHODS: In a multicentre, open-label, exploratory, non-interventional study, inhaled tobramycin (300 mg twice daily) and colistin (1 million I.U. twice daily) were sequentially combined with the aim to investigate the effect on 41 CF patients with chronic P. aeruginosa infections for six months (mean age 24 ± 10.8y). RESULTS: Six patients had adverse events that were assessed as being related to treatment. Mucus production and coughing both decreased in 39%, whereas FEV1 absolute and relative to baseline increased by 4.9% and 9.1%, respectively (p = 0.004) in 29 patients, who were definitely treated sequentially. Efficacy of the therapy was rated 'excellent' or 'good' by the physicians in 80.5% of the patients. CONCLUSIONS: The results indicate that treatment with inhaled antibiotics, sequentially combined, was very well tolerated by most patients and may have a beneficial effect, even if transitory on lung function and respiratory symptoms.
Asunto(s)
Antibacterianos/uso terapéutico , Colistina/uso terapéutico , Fibrosis Quística/tratamiento farmacológico , Infecciones por Pseudomonas/tratamiento farmacológico , Tobramicina/uso terapéutico , Administración por Inhalación , Adolescente , Adulto , Enfermedad Crónica , Fibrosis Quística/microbiología , Fibrosis Quística/patología , Esquema de Medicación , Combinación de Medicamentos , Femenino , Volumen Espiratorio Forzado/efectos de los fármacos , Humanos , Masculino , Estudios Prospectivos , Infecciones por Pseudomonas/microbiología , Infecciones por Pseudomonas/patología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/crecimiento & desarrollo , Resultado del TratamientoAsunto(s)
Asma , Regulación de la Expresión Génica/inmunología , Interleucina-9 , Factores de Transcripción NFATC , Asma/inmunología , Asma/metabolismo , Niño , Preescolar , Femenino , Humanos , Lactante , Interleucina-9/inmunología , Interleucina-9/metabolismo , Masculino , Factores de Transcripción NFATC/biosíntesis , Factores de Transcripción NFATC/inmunologíaRESUMEN
Males with deletions of or within Xp22.3-pter display variable contiguous gene syndromes including manifestations of Léri-Weill syndrome, chondrodysplasia punctata, mental retardation, ichthyosis, Kallmann syndrome, and ocular albinism. Herein, we report on a male infant with a large, cytogenetically visible, terminal Xp deletion defined by extensive FISH and STS marker analysis to encompass 9.6 Mb, and findings of all of the disorders mentioned above. His deletion approximates the largest Xp terminal deletion ever reported in a male individual. Since the extent of terminal Xp deletions viable in males is limited by the position of male lethal genes in Xp22.2 at about 10-11 Mb from the telomere, this patient falls into the category of the most severe male terminal Xp deletion phenotype.
Asunto(s)
Anomalías Múltiples/genética , Anomalías Múltiples/patología , Deleción Cromosómica , Cromosomas Humanos X/genética , Albinismo Ocular/genética , Albinismo Ocular/patología , Condrodisplasia Punctata/genética , Condrodisplasia Punctata/patología , Humanos , Ictiosis/genética , Ictiosis/patología , Hibridación Fluorescente in Situ , Lactante , Discapacidad Intelectual/genética , Discapacidad Intelectual/patología , Síndrome de Kallmann/genética , Síndrome de Kallmann/patología , MasculinoRESUMEN
Atherosclerosis is associated with alterations in nitric oxide (NO)/cGMP signaling. In early stages of the disease, inflammatory and possibly other cells produce reactive oxygen species that scavenge vasoprotective NO. In addition to the oxidative stress, expression and activity of enzymes downstream to NO formation may also be affected. Here, we show in the aortas of chronically hypercholesterolemic rabbits (a model of late-stage atherosclerosis), both subunits and specific activity of the NO receptor soluble guanylyl cyclase (sGC) were significantly reduced, whereas overall NO synthase activity was unaffected. These changes were most prominent in the neointimal layer, wherein cGMP-dependent protein kinase I (cGK) levels also were reduced. Additionally, a protein (p38(nt)) that was constitutively tyrosine-nitrated was detected, and its expression was significantly reduced in atherosclerotic aorta. Phosphorylation of the cGK substrate vasodilator-stimulated phosphoprotein (VASP) at Ser-239, an established biochemical endpoint of NO/cGMP signaling, also was reduced. Thus, late-stage atherosclerosis is associated not only with enhanced NO breakdown but also with altered NO reception and cGMP signaling. Preferential down-regulation in neointima suggests a direct connection of these changes to neointimal proliferation and vascular dysfunction and provides a rationale for future pharmacotherapy using classical and novel sGC activators.
Asunto(s)
Arteriosclerosis/patología , Arteriosclerosis/fisiopatología , GMP Cíclico/fisiología , Animales , Aorta Torácica/patología , Aorta Torácica/fisiopatología , Moléculas de Adhesión Celular/química , Moléculas de Adhesión Celular/fisiología , Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Guanilato Ciclasa/metabolismo , Lípidos/sangre , Proteínas de Microfilamentos , Óxido Nítrico/fisiología , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico Sintasa de Tipo II , Ácido Peroxinitroso/metabolismo , Fosfoproteínas/química , Fosfoproteínas/fisiología , Hidrolasas Diéster Fosfóricas/metabolismo , Fosforilación , Conejos , Transducción de Señal , Superóxido Dismutasa/metabolismoRESUMEN
The endothelium-derived relaxing factors nitric oxide (NO) and prostacyclin (PGI(2)) are important antithrombotic, relaxant, and antiproliferative agents of the blood vessel wall that exert their intracellular effects primarily via cGMP- and cAMP-dependent protein kinases (cGK, cAK). However, no biochemical marker for their activity in the intact blood vessel is available except for transient increases in the concentration of cGMP and cAMP. Using Western blot analysis and specific antibodies, we show here that phosphorylation of the vasodilator-stimulated phosphoprotein (VASP) at Ser239 (P(Ser239)-VASP) in rabbit aorta was detectable only in segments with an intact endothelium, although at least one third of VASP is contained in the remaining vascular wall. In endothelium-denuded aorta, VASP phosphorylation was increased by the NO donor sodium nitroprusside (SNP). Levels of P(Ser239)-VASP, in the presence of endothelium and either SNP or 8-bromo-cAMP, were maximal. VASP phosphorylation elicited by 8-bromo-cAMP was inhibited significantly by the cGK inhibitor Rp-8-Br-PET-cGMPS. Stimulated P(Ser239)-VASP formation was fully reversible, reaching basal levels after 10 min of repeated washouts. Consistent with the important role that the NO/cGMP pathway plays in the formation of P(Ser239)-VASP in rabbit aorta, inhibition of NO synthase by N(omega)-nitro-L-arginine methyl ester (L-NAME; 1 mM) or of soluble guanylyl cyclase by 1H-[1,2,4]oxadiazolo[3,4-a]quinoxalin-1-one (ODQ; 50 microM) almost completely abolished P(Ser239)-VASP formation in endothelium intact blood vessels. These data suggest that vascular P(Ser239)-VASP is primarily regulated by the NO/cGMP pathway and may thus serve as a biochemical marker for the activity state of this essential pathway in endothelial function.