ABSTRACT
Idiopathic pulmonary hypertension (IPAH) is a rare disease characterized by a progressive increase in pulmonary vascular resistance leading to heart failure. MicroRNAs (miRNAs) are small noncoding RNAs that control the expression of genes, including some involved in the progression of IPAH, as studied in animals and lung tissue. These molecules circulate freely in the blood and their expression is associated with the progression of different vascular pathologies. Here, we studied the expression profile of circulating miRNAs in 12 well-characterized IPAH patients using microarrays. We found significant changes in 61 miRNAs, of which the expression of miR23a was correlated with the patients' pulmonary function. We also studied the expression profile of circulating messenger RNA (mRNAs) and found that miR23a controlled 17% of the significantly changed mRNA, including PGC1α, which was recently associated with the progression of IPAH. Finally we found that silencing of miR23a resulted in an increase of the expression of PGC1α, as well as in its well-known regulated genes CYC, SOD, NRF2, and HO1. The results point to the utility of circulating miRNA expression as a biomarker of disease progression.
Subject(s)
Familial Primary Pulmonary Hypertension/genetics , MicroRNAs/metabolism , Adult , Aged , Biomarkers/metabolism , Cells, Cultured , Cytochromes c/genetics , Cytochromes c/metabolism , Familial Primary Pulmonary Hypertension/pathology , Female , Gene Expression Profiling , Heme Oxygenase-1/genetics , Heme Oxygenase-1/metabolism , Humans , Male , Middle Aged , NF-E2-Related Factor 2/genetics , NF-E2-Related Factor 2/metabolism , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolism , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
Introducción: El humo de tabaco es la principal causa de la inflamación en la EPOC. Los mecanismos quediferencian a los fumadores que desarrollan EPOC son diversos. En este estudio analizamos la diferentepresencia de citocinas en secreciones respiratorias de pacientes fumadores con o sin EPOC y las propiedadessecretoras del epitelio bronquial diferenciado, obtenido de los propios individuos tras su exposiciónal humo de tabaco.Material y métodos: Se estudió a 27 pacientes fumadores, 12 de ellos con EPOC no tratados previamentecon esteroides. En 11 se obtuvo la muestra mediante esputo inducido y el resto procedía del aspiradobronquial tras fibrobroncoscopia. Se determinaron las concentraciones de IL8, IL13 y TNF en el sobrenadante.Se compararon los resultados obtenidos entre individuos con o sin EPOC y se investigó su relacióncon la gravedad de la EPOC expresada según el grado de obstrucción, disnea, presencia de hipersecrecióne intensidad del tabaquismo. Se obtuvieron cultivos de células diferenciadas epiteliales bronquiales,mediante interfase aire-líquido en 4 individuos fumadores. Las muestras fueron expuestas a concentracionescrecientes de humo de tabaco (5-20%) y se determinó la expresión epitelial de ARNm de Muc5AC,IL8 y TNF .Resultados: Los pacientes con EPOC tenían valores significativamente más altos de IL8 que los fumadoressanos (41 [22] vs. 21 [12] pM). Los valores de IL8 se correlacionaron de forma significativa con la gravedadde la obstrucción (r = 0,6; p < 0,05), disnea (r = 0,45; p < 0,05) y la presencia de hipersecreción. No habíarelación entre las citocinas y la intensidad o duración del hábito tabáquico. El humo de tabaco produjoun incremento dependiente de la dosis en la expresión de ARNm para Muc5AC, IL8 y TNF. Conclusiones: Existen diferencias en la producción de citocinas, fundamentalmente IL8, entre individuosfumadores sanos o con EPOC que podrían ser justificadas por la acción directa del humo de tabaco sobrelas células epiteliales (AU)
Introduction: Cigarette smoke is the main cause of inflammation in COPD. The mechanisms that differentiatesmokers who develop COPD are diverse. In this study, we analyzed the presence of cytokines in therespiratory secretions of smokers with or without COPD and the secretory properties of the differentiatedbronchial epithelium obtained from the individuals themselves after exposure to tobacco smoke.Material and methods: Twenty-seven smokers were studied, 12 of whom had COPD that had not beenpreviously treated with steroids. In 11, samples were obtained by means of induced sputum, and the remaining samples were collected from bronchial aspiration after bronchoscopy. Concentrations of IL-8,IL-13 and TNF in the supernatant were determined. The results obtained were compared between individualswith and without COPD, and we studied their relationship with the severity of COPD as expressedby the degree of obstruction, dyspnea, presence of hypersecretion and intensity of smoking. Bronchialepithelial cell cultures were obtained by air-liquid interface in 4 smokers. The samples were exposed toincreasing concentrations of cigarette smoke (5-20%) and the epithelial mRNA expressions of MUC5AC,IL8 and TNF were determined.Results: COPDpatients had significantly higher values of IL-8 than healthy smokers (41 [22] vs. 21 [12] pM).The values of IL-8 correlated significantly with the severity of the obstruction (r = 0.6; p < 0.05), dyspnea(r = 0.45; p < 0.05) and the presence of hypersecretion. There was no relationship between cytokines andthe intensity or duration of the tobacco habit. Cigarette smoke produced a dose-dependent increase inthe expression of RNAm for Muc5AC, IL8 and TNF .Conclusions: There are differences in cytokine production (fundamentally IL8) between smokers and smokerswith COPD which could be explained by the direct action of cigarette smoke on epithelial cells (AU)
Subject(s)
Humans , Male , Female , Young Adult , Adult , Middle Aged , Aged , Pulmonary Disease, Chronic Obstructive/etiology , Smoking/adverse effects , Respiratory Mucosa/pathology , Interleukin-8/biosynthesis , Tumor Necrosis Factor-alpha/biosynthesis , Inflammation , Interleukin-13/biosynthesis , Sputum/chemistry , Mucin 5AC/biosynthesisABSTRACT
El roflumilast (3-ciclopropilmetoxi-4-difluorometoxi-N-[3,5-di-cloro-4-piridil]-benzamida) ha sido el primeragente de una nueva clase farmacológica, los inhibidores selectivos de la fosfodiesterasa-4 (IPDE4), aprobadopara el tratamiento de los pacientes con enfermedad pulmonar obstructiva crónica (EPOC). El mecanismomolecular de acción del roflumilast es la inhibición de la isoenzima de la PDE4 y el consecuente incrementodel monofosfato de adenosina cíclico. Es evidente que el roflumilast muestra varios efectos farmacológicos:antiinflamatorio, antienfisematoso, antifibrótico, inhibidor de la hipertensión pulmonar y mucorregulador.Las acciones farmacológicas responsables de sus efectos son: a) la inhibición de la formación de especiesreactivas de oxígeno en las células del epitelio, los neutrófilos y las células del músculo liso; b) la inhibición dela proliferación de las células del músculo liso de la arteria pulmonar, las células endoteliales y, probablemente,algunas células inflamatorias responsables del remodelado vascular pulmonar; c) la inhibición de los fibroblastos,con la consiguiente disminución del remodelado pulmonar, y, finalmente, d) la inhibición de la producciónde moco y la mejora del batido ciliar.En suma, el roflumilast es el primer antiinflamatorio no esteroideo que podrá utilizarse en el tratamiento dela EPOC(AU)
Roflumilast (3-cyclopropylmethoxy-4-difluoromethoxy-n-(3,5-dichloropyrid-4-yl)benzamide) was the firstagent of a novel pharmacological class, selective phosphodiesterase 4 (PDE4) inhibitors, approved for the use ofchronic obstructive pulmonary disease (COPD). The molecular mechanism of action of roflumilast is inhibitionof the PDE4 isoenzyme with a consequent increase of cyclic adenosine monophosphate. Roflumilast evidentlyhas several pharmacological effects: antiinflammatory, anti-emphysema, and antibiotic actions. This drugalso inhibits pulmonary hypertension and reduces mucus hypersecretion. The pharmacological actionsleading to these effects are: a) inhibition of reactive oxygen species formation in epithelial cells, neutrophilsand smooth muscle cells; b) inhibition of smooth muscle cell proliferation in the pulmonary artery, endothelialcells and probably some inflammatory cells causing pulmonary vascular remodeling; c) inhibition offibroblasts, with a consequent reduction in pulmonary remodeling and, finally, d) inhibition of mucusproduction and improved ciliary beat frequency. In summary, roflumilast is the first non-steroidalanti-inflammatory drug that can be used in the treatment of COPD(AU)
Subject(s)
Humans , Pulmonary Disease, Chronic Obstructive/drug therapy , Phosphodiesterase Inhibitors/pharmacokinetics , Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Pulmonary Emphysema/drug therapySubject(s)
Humans , Phosphodiesterase Inhibitors/pharmacokinetics , Pulmonary Disease, Chronic Obstructive/drug therapy , Inflammation/drug therapy , Nucleotides, Cyclic/pharmacokinetics , Phosphoric Diester Hydrolases , Isoenzymes , Pulmonary Disease, Chronic Obstructive/physiopathology , Anti-Inflammatory Agents/pharmacokineticsABSTRACT
La discinesia ciliar primaria es un trastorno genéticamente determinado que se caracteriza por un movimiento ciliar alterado o ausente. Genera un déficit en el aclaramiento mucociliar que se manifiesta clínicamente como infecciones crónicas de vías aéreas constantes desde el nacimiento, así como esterilidad masculina por inmovilidad del espermatozoide y situs inversus en el 4050% de los pacientes (síndrome de Kartagener). El diagnóstico se basa en el estudio de la movilidad ciliar mediante vídeo de alta resolución digital y alta velocidad, complementado con el estudio de la ultraestructura ciliar. La amplia distribución ciliar en el organismo y sus numerosas funciones hacen que su patología origine, además de la discinesia ciliar primaria, otras ciliopatías (AU)
Primary ciliary dyskinesia is a genetically inherited syndrome characterized by cilia immotility or dysmotility. Deficiency in mucociliary clearance produces chronic respiratory infections since birth, male sterility by spermatozoid immotility and situs inversus in 4050% of patients (Kartagener's syndrome). Diagnosis is made by analyzing cilia motility with high-speed digital video and ciliar ultrastructure. The wide distribution and functions of the cilia in the body mean that this dysfunction can generate other ciliopathies apart from primary ciliary dyskinesia (AU)
Subject(s)
Humans , Male , Female , Kartagener Syndrome/epidemiology , Sinusitis/complications , Bronchiectasis/complications , Situs Inversus/complications , Ciliary Motility Disorders/therapy , Ciliary Motility Disorders/diagnosis , Diagnosis, Differential , Organelles/genetics , Organelles/pathologyABSTRACT
Roflumilast (3-cyclopropylmethoxy-4-difluoromethoxy-n-(3,5-dichloropyrid-4-yl)benzamide) was the first agent of a novel pharmacological class, selective phosphodiesterase 4 (PDE(4)) inhibitors, approved for the use of chronic obstructive pulmonary disease (COPD). The molecular mechanism of action of roflumilast is inhibition of the PDE(4) isoenzyme with a consequent increase of cyclic adenosine monophosphate. Roflumilast evidently has several pharmacological effects: antiinflammatory, anti-emphysema, and antibiotic actions. This drug also inhibits pulmonary hypertension and reduces mucus hypersecretion. The pharmacological actions leading to these effects are: a) inhibition of reactive oxygen species formation in epithelial cells, neutrophils and smooth muscle cells; b) inhibition of smooth muscle cell proliferation in the pulmonary artery, endothelial cells and probably some inflammatory cells causing pulmonary vascular remodeling; c) inhibition of fibroblasts, with a consequent reduction in pulmonary remodeling and, finally, d) inhibition of mucus production and improved ciliary beat frequency. In summary, roflumilast is the first non-steroidal anti-inflammatory drug that can be used in the treatment of COPD.
Subject(s)
Aminopyridines/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Benzamides/pharmacology , Phosphodiesterase 4 Inhibitors/pharmacology , Airway Remodeling/drug effects , Aminopyridines/pharmacokinetics , Aminopyridines/therapeutic use , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Benzamides/pharmacokinetics , Benzamides/therapeutic use , Cell Division/drug effects , Cilia/drug effects , Cyclopropanes/pharmacokinetics , Cyclopropanes/pharmacology , Cyclopropanes/therapeutic use , Drug Evaluation, Preclinical , Fibroblasts/drug effects , Humans , Hypertension, Pulmonary/drug therapy , Macrophages/drug effects , Mice , Molecular Structure , Mucus/metabolism , Myocytes, Smooth Muscle/drug effects , Neutrophils/drug effects , Oxidants/toxicity , Oxidative Stress/drug effects , Phosphodiesterase 4 Inhibitors/pharmacokinetics , Phosphodiesterase 4 Inhibitors/therapeutic use , Pulmonary Disease, Chronic Obstructive/drug therapy , Pulmonary Emphysema/drug therapy , Rats , Smoke/adverse effects , NicotianaSubject(s)
Anti-Inflammatory Agents/therapeutic use , Phosphodiesterase 4 Inhibitors/therapeutic use , Pulmonary Disease, Chronic Obstructive/drug therapy , Pulmonary Emphysema/drug therapy , Aminopyridines/pharmacokinetics , Aminopyridines/therapeutic use , Benzamides/pharmacokinetics , Benzamides/therapeutic use , Cilia/drug effects , Cilia/physiology , Cyclic AMP/physiology , Cyclic GMP/physiology , Cyclopropanes/pharmacokinetics , Cyclopropanes/therapeutic use , Humans , Inflammation , Isoenzymes/physiology , Mucins/metabolism , Oxidative Stress/drug effects , Phosphoric Diester Hydrolases/physiology , Pulmonary Circulation/drug effects , Pulmonary Disease, Chronic Obstructive/enzymology , Pulmonary Emphysema/enzymology , Second Messenger SystemsABSTRACT
Primary ciliary dyskinesia is a genetically inherited syndrome characterized by cilia immotility or dysmotility. Deficiency in mucociliary clearance produces chronic respiratory infections since birth, male sterility by spermatozoid immotility and situs inversus in 40-50% of patients (Kartagener's syndrome). Diagnosis is made by analyzing cilia motility with high-speed digital video and ciliar ultrastructure. The wide distribution and functions of the cilia in the body mean that this dysfunction can generate other ciliopathies apart from primary ciliary dyskinesia.
Subject(s)
Ciliary Motility Disorders , Ciliary Motility Disorders/diagnosis , Ciliary Motility Disorders/therapy , HumansABSTRACT
La identificación de la fosfodiesterasa de nucleótidos cíclicos (PDE), enzima responsable de la destrucción del AMPc y el GMPc intracelular, como punto de ación para las metilxantinas ha originado una creciente actividad investigadora en este campo. Esto ha tenido como resultado la caracterización de múltiples isoenzimas de PDE (PDE 1 a PDE 9), su distribución tisular específica, y el descubrimiento y desarrollo de fármacos inhibidores selectivos para algunosde estos isoenzimas. La disponibilidad de los fármacos inhibidores selectivos de isoenzimas de PDE ha permitido estudios experimentales in vitro e in vivo, cuyo fin era determinar su valor potencial como fármacos antiasmáticos. Aunque la investigación básica está siendo muy importante, la mayoría de los fármacos inhibidores selectivos de PDE están empezando a someterse a ensayos clínicos para valorar su utilidad en el tratamiento de esta patología. La investigación futura debe dirigirse a conocer mejor la distribución tisular de la PDE y su papel en la fisiopatología, así como para desarrollar mejores (más selectivas) moléculas inhibidoras de la fosfodiesterasa. (AU)
