Nuevos fármacos antiasmáticos : inhibición selectiva de isoenzimas de la fosfodiesterasa

The identification of cyclic nucleotide phosphodiesterase (PDE), the enzyme responsible for the intracellular degradation of cAMP and cGMP, as the target for methyxanthines has given rise to a reserch effort resulting in the characterization of multiple PDE isoenzymes (PDE 1 to PDE 9), their specific tissular distribution and development of selective inhibitors for some of these isoenzymes. This bioavailability of theese selective PDE isoenzyme inhibitors has permitted studies with regard to their potential value as antiasthmatic drugs. Although the basic research is being intensive, most of the selective PDE isoenzyme inhibitors are beginning to be subjected to clinical trials to asses their usefulness in the treatment of this pathology. Future research should be aimed at ascertaining the tissular distribution of the PDEs and their role in physiophathology, phosphodiesterase inhibitors.La identificación de la fosfodiesterasa de nucleótidos cíclicos (PDE), enzima responsable de la destrucción del AMPc y el GMPc intracelular, como punto de acción para las metilxantinas ha originado una creciente actividad investigadora en este campo. Esto ha tenido como resultado la caracterización de múltiples isoenzimas de PDE (PDE1 a PDE 9), su distribución tisular específica, y el descubrimiento y desarrollo de fármacos inhibidores selectivos para algunos de estos iosenzimas. 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 paramoléculas inhibidoras de la fosfodiesterasa

Roflumilast N-oxide inhibits bronchial epithelial to mesenchymal transition induced by cigarette smoke in smokers with COPD

BACKGROUND: Epithelial to mesenchymal transition (EMT) is under discussion as a potential mechanism of small airway remodelling in COPD. In bronchial epithelium of COPD and smokers markers of EMT were described. In vitro, EMT may be reproduced by exposing well-differentiated human bronchial epithelial cells (WD-HBEC) to cigarette smoke extract (CSE). EMT may be mitigated by an increase in cellular cAMP. OBJECTIVE: This study explored the effects of roflumilast N-oxide, a PDE4 inhibitor on CSE-induced EMT in WD-HBEC and in primary bronchial epithelial cells from smokers and COPD in vitro. METHODS: WD-HBEC from normal donors were stimulated with CSE (2.5%) for 72 h in presence of roflumilast N-oxide (2 nM or 1 μM) or vehicle. mRNA and protein of EMT markers αSMA, vimentin, collagen-1, E-cadherin, ZO-1, KRT5 as well as NOX4 were quantified by real-time quantitative PCR or protein array, respectively. Phosphorylated and total ERK1/2 and Smad3 were assessed by protein array. cAMP and TGFβ1 were measured by ELISA. Reactive oxygen species (ROS) were determined by DCF fluorescence, after 30 min CSE (2.5%). Apoptosis was measured with Annexin V/PI labelling. In some experiments, EMT markers were determined in monolayers of bronchial epithelial cells from smokers, COPD versus controls. RESULTS: Roflumilast N-oxide protected from CSE-induced EMT in WD-HBEC. The PDE4 inhibitor reversed both the increase in mesenchymal and the loss in epithelial EMT markers. Roflumilast N-oxide restored the loss in cellular cAMP following CSE, reduced ROS, NOX4 expression, the increase in TGFβ1 release, phospho ERK1/2 and Smad3. The PDE4 inhibitor partly protected from the increment in apoptosis with CSE. Finally the PDE4 inhibitor decreased mesenchymal yet increased epithelial phenotype markers in HBEC of COPD and smokers. CONCLUSIONS: Roflumilast N-oxide may mitigate epithelial-mesenchymal transition in bronchial epithelial cells in vitro

Molecular and Clinical Predictors of Quality of Life in Chronic Rhinosinusitis with Nasal Polyps

Factors influencing the quality of life (QoL) of patients with chronic rhinosinusitis with nasal polyposis (CRSwNP) are poorly understood. We set out to determine the predictive factors on patients' QoL using the Sino-Nasal Outcome Test-22 (SNOT-22); (2) Methods: An ambispective analysis of data from patients diagnosed with CRSwNP in our institution. All the patients underwent a nasal polyp biopsy and completed the SNOT-22 questionnaire. Demographic and molecular data as well as the SNOT-22 scores were collected. Patients were classified in six subgroups considering the presence of asthma, non-steroidal drugs (NSAID) intolerance and corticosteroid resistance; (3) Results: The mean SNOT-22 score was 39. Considering the clinical parameters, the SNOT-22 value was significantly associated with NSAID intolerance (p = 0.04) and the endoscopic polyp score (p = 0.04). A high SNOT-22 value was also correlated with high tissue eosinophilia (p = 0.01) and high IL-8 expression; (4) Conclusions: Eosinophilia, IL-8 expression and NSAID intolerance can be used as predictors of worse QoL in patients with CRSwNP

Angiotensin II Induces Neutrophil Accumulation In Vivo Through Generation and Release of CXC Chemokines

Background— Angiotensin II (Ang II) is implicated in the development of cardiac ischemic disorders in which prominent neutrophil accumulation occurs. Ang II can be generated intravascularly by the renin-angiotensin system or extravascularly by mast cell chymase. In this study, we characterized the ability of Ang II to induce neutrophil accumulation. Methods and Results— Intraperitoneal administration of Ang II (1 nmol/L) induced significant neutrophil recruitment within 4 hours (13.3±2.3x106 neutrophils per rat versus 0.7±0.5x106 in control animals), which disappeared by 24 hours. Maximal levels of CXC chemokines were detected 1 hour after Ang II injection (577±224 pmol/L cytokine-inducible neutrophil chemoattractant [CINC]/keratinocyte-derived chemokine [KC] versus 5±3, and 281±120 pmol/L macrophage inflammatory protein [MIP-2] versus 14±6). Intravital microscopy within the rat mesenteric microcirculation showed that the short-term (30 to 60 minutes) leukocyte–endothelial cell interactions induced by Ang II were attenuated by an anti-rat CINC/KC antibody and nearly abolished by the CXCR2 antagonist SB-517785-M. In human umbilical vein endothelial cells (HUVECs) or human pulmonary artery media in culture, Ang II induced interleukin (IL)-8 mRNA expression at 1, 4, and 24 hours and the release of IL-8 at 4 hours through interaction with Ang II type 1 receptors. When HUVECs were pretreated with IL-1 for 24 hours to promote IL-8 storage in Weibel-Palade bodies, the Ang II–induced IL-8 release was more rapid and of greater magnitude. Conclusions— Ang II provokes rapid neutrophil recruitment, mediated through the release of CXC chemokines such as CINC/KC and MIP-2 in rats and IL-8 in humans, and may contribute to the infiltration of neutrophils observed in acute myocardial infarction.Mata Roig, Manuel, [email protected] ; Cortijo Gimeno, Julio, [email protected] ; Morcillo Sanchez, Esteban Jesus, [email protected] ; Jose, Peter J., [email protected] ; Sanz Ferrando, Maria Jesus, [email protected]

Modificación de la morfología mitocondrial en ventrículo aislado, en relación con el tiempo de actividad funcional

Se estudia mediante microscopía electrónica, las alteraciones de las mitocondrias en el ventrículo aislado de rata estimulado eléctricamente y registrándose isométricamente la contracción. Se observa una disminución progresiva de la fuerza contráctil (27,94%, n=6). Port y col. relacionan la disminución funcional con el envejecimiento del órgano. Se compara el estado de las mitcondrias al comienzo y al final del experimento (3 horas), observándose la desintegración de las crestas mitocondriales. Estos resultados se complementan con los de otros autores referentes al acúmulo de grasa y disminución del retículo sarcoplásmico. Pensamos continuar estos estudios sobre el 'envejecimiento' de los órganos aislados, dada la importancia que tiene en la investigación farmacológica. Departamento de Famacología y Farmacotecnia. Facultat de Medicina i Odontologia de València

Role of Circulating miRNAs as Biomarkers in Idiopathic Pulmonary Arterial Hypertension: Possible Relevance of miR-23a

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

Role of Circulating miRNAs as Biomarkers in Idiopathic Pulmonary Arterial Hypertension: Possible Relevance of miR-23a

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 PGC1 , which was recently associated with the progression of IPAH. Finally we found that silencing of miR23a resulted in an increase of the expression of PGC1 , 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

High-Speed Video Microscopy For Primary Ciliary Dyskinesia Diagnosis: A Study of Ciliary Motility Variations with Time and Temperature

Primary ciliary dyskinesia (PCD) is a rare disease resulting from a defect in ciliary function that generates, among other issues, chronic upper and lower respiratory tract infections. European guidelines recommend studying ciliary function (pattern (CBP) and frequency (CBF)), together with characteristic clinical symptoms, as one of the definitive tests. However, there is no "gold standard". The present study aims to use high-speed video microscopy to describe how CBF and CBP alter over time and at different temperatures to reduce the error rate in the diagnosis of PCD. Samples of nasal epithelium from 27 healthy volunteers were studied to assess CBF and CBP at 0, 3, 24, 48, and 72 h, at room temperature and 4 ◦C. It was observed that CBF increased while CBP became dyskinetic, both at room temperature and at 4 ◦C, as time passed, especially after 3 h. In order to preserve all ciliary function parameters and to perform a reliable analysis to improve the diagnostic process of PCD, analysis should be performed within the first 3 h of sample collection, preferably in reference center