RESUMEN
TGF-beta-targeting structural and inflammatory cells has been implicated in the mechanisms leading to the inflammatory and restructuring processes in asthma, suggesting an impact of TGF-beta1 signaling on the development and persistency of this disease. We investigated the potential early involvement of TGF-beta1 activity in the immunological and molecular mechanisms underlying progression of inflammation in childhood asthma. We evaluated the levels of TGF-beta1 in induced sputum supernatants (ISSs) and the expression of small mother cell against decapentaplegic (Smad) 2 and Smad7 proteins in induced sputum cells (ISCs) from children with intermittent asthma (IA), moderate asthma (MA) and control subjects (C). Furthermore, we investigated the regulatory role of TGF-beta1 activity on eosinophil and neutrophil adhesion to epithelial cells using adhesion assay, and on the granulocyte expression of adhesion molecule CD11b/CD18 Macrophage-1 antigen (MAC-1), by flow cytometry. We found that the levels of TGF-beta1 are increased in ISSs of IA and MA in comparison to C, concomitantly to the activation of intracellular signaling TGFbeta/Smads pathway in ISCs. In MA, TGF-beta1 levels correlated with the number of sputum eosinophils and neutrophils. Furthermore, we showed the ability of sputum TGF-beta1 to promote eosinophil and neutrophil adhesion to epithelial cells, and to increase the expression of MAC-1 on the granulocyte surface. This study shows the activation of TGFbeta/Smad signaling pathway in the airways of children with IA and, despite the regular ICS treatment, in children with MA, and provides evidence for the contribution of TGF-beta1 in the regulation of granulocyte activation and trafficking.
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Asma/metabolismo , Pulmón/metabolismo , Transducción de Señal , Factor de Crecimiento Transformador beta1/metabolismo , Administración por Inhalación , Adolescente , Corticoesteroides/administración & dosificación , Factores de Edad , Asma/diagnóstico , Asma/tratamiento farmacológico , Asma/inmunología , Asma/fisiopatología , Broncodilatadores/administración & dosificación , Estudios de Casos y Controles , Adhesión Celular , Línea Celular , Niño , Eosinófilos/inmunología , Eosinófilos/metabolismo , Células Epiteliales/inmunología , Células Epiteliales/metabolismo , Femenino , Granulocitos/inmunología , Granulocitos/metabolismo , Humanos , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/fisiopatología , Antígeno de Macrófago-1/metabolismo , Masculino , Neutrófilos/inmunología , Neutrófilos/metabolismo , Fosforilación , Mucosa Respiratoria/inmunología , Mucosa Respiratoria/metabolismo , Índice de Severidad de la Enfermedad , Transducción de Señal/efectos de los fármacos , Proteína Smad2/metabolismo , Proteína smad7/metabolismo , Esputo/metabolismoRESUMEN
United airway disease (UAD) concept proposed that asthma and rhinitis are both different clinical manifestation of a single inflammatory process. The aim of this study is to assess in upper and lower airways the level of inflammation and oxidative stress and to investigate the relationship between biomarkers in persistent allergic rhinitis (PER) and in concomitant asthma with PER. By a crosssectional study we measured oral and nasal (FENO) and oral and nasal EBC 8-isoprostane, LTB4 and PGE2 in children with PER (n=14) and with PER and concomitant intermittent asthma (IA; n=25), mild persistent asthma (mA; n=28), moderate persistent asthma (MA; n=13) and in Healthy Controls (HCs; n=13). Oral and nasal FENO concentrations were increased in children with PER, IA, mA and MA when compared with HCs. Nasal 8-isoprostane was higher in EBC of children with PER and asthma than in HCs. Oral and nasal LTB4 were higher in EBC of children with PER and mA than in HCs. Oral and nasal PGE2 concentrations were higher in EBC of children with PER than in HCs. Positive correlations between oral and nasal biomarkers were found in IA for LTB4 and PGE2, in mA for FENO, 8-isoprostane, LTB4 and PGE2, and in MA for PGE2. No correlations were observed in children with PER and HCs. Our results suggest that non-invasive markers of inflammation and oxidative stress might be useful to study the relationships between oral and nasal compartments in allergic children with PER and concomitant asthma with the aim of defining the UAD.
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Asma/metabolismo , Inflamación/diagnóstico , Mucosa Bucal/metabolismo , Mucosa Nasal/metabolismo , Estrés Oxidativo , Rinitis Alérgica Perenne/metabolismo , Adolescente , Pruebas Respiratorias , Niño , Estudios Transversales , Dinoprostona/análisis , Femenino , Humanos , Leucotrieno B4/análisis , Masculino , Óxido Nítrico/metabolismoRESUMEN
BACKGROUND AND AIM: The short, repetitive hypoxaemic episodes observed in obstructive sleep apnoea (OSA) may determine small augmentations in mature red blood cells. It is unknown whether they affect reticulocyte release. This study explored whether the number and degree of maturation of circulating reticulocytes may be altered in OSA, possibly through the effect of erythropoietin. METHODS: Fifty male adult patients with suspected OSA, normoxic during wakefulness, were studied. After nocturnal polysomnography, a blood sample was withdrawn for blood cells count, erythropoietin, iron and transferrin determination. Reticulocyte concentration and degree of immaturity [high (H), medium (M), or low (L)] were also determined. Immature reticulocyte fraction (IRF) was calculated as (M+H) percentage of reticulocytes. RESULTS: A wide range of OSA severity was found [apnoea/hypopnoea index (AHI): 44.3 +/- 30.4, range 0.3-105; sleep time spent at oxyhaemoglobin saturation <90%: 18.1 +/- 22.2%, range 0-81%]. Both reticulocyte count and IRF slightly exceeded the normal range. Patients with a reticulocyte concentration > 2% had higher EPO levels (p < 0.05), but not worse nocturnal desaturations, than those with values < 2%. By contrast, subjects with IRF < 15% showed worse desaturations (p < 0.05), but similar EPO concentrations, when compared to subjects whose IRF was < 10%. At univariate analysis, reticulocyte count correlated to erythropoietin, while IRF to transferrin saturation, BMI and OSA severity. At multiple regression, only lowest nocturnal oxygen saturation remained a significant contributor to IRF (r2 0.223, p < 0.05). CONCLUSIONS: This data suggests that hypoxaemia due to OSA could influence the release of immature reticulocytes, but this effect is not mediated by erythropoietin.
Asunto(s)
Recuento de Reticulocitos , Apnea Obstructiva del Sueño/sangre , Adulto , Estudios de Cohortes , Eritropoyesis/fisiología , Eritropoyetina/sangre , Humanos , Masculino , Persona de Mediana Edad , Polisomnografía , Índice de Severidad de la Enfermedad , Apnea Obstructiva del Sueño/complicaciones , Transferrina/metabolismoRESUMEN
In asthma a dysregulation of eosinophil apoptosis and an imbalance of metalloproteinase-9 (MMP-9) and tissue inhibitor metalloproteinase-1 (TIMP-1) play an important role in airway inflammation and remodelling. We evaluated the effects of a low dose of inhaled fluticasone proprionate (FP) (100 microg bid by Diskus) for 4 weeks in 24 steroid naive patients with mild persistent asthma, symptomatic and with a sputum eosinophilia >or=3% on clinical outcomes and inflammatory markers such as the induced sputum eosinophils, the induced sputum apoptotic eosinophils, the levels of MMP-9 and TIMP-1 and their molar ratio in the induced sputum supernatants. After FP treatment forced expiratory volume (FEV1) and FEV1/forced vital capacity values, PEF (L/min), sputum apoptotic eosinophils, and MMP-9/TIMP-1 molar ratio in sputum supernatants of asthmatic subjects were significantly increased in comparison with baseline, while sputum eosinophils significantly decreased. Change (Delta) in FEV1 after treatment with FP negatively correlated with the Delta in sputum eosinophils, while the Delta in MMP-9 values positively correlated with Delta in TIMP-1 values. This study shows that the clinical improvement achieved by the use of low doses of FP in asthmatics is related, at least in part, to the resolution of eosinophilic inflammation and the downregulation of remodelling markers.
Asunto(s)
Androstadienos/administración & dosificación , Antiinflamatorios/administración & dosificación , Asma/tratamiento farmacológico , Asma/fisiopatología , Inflamación/tratamiento farmacológico , Metaloproteinasa 9 de la Matriz , Administración por Inhalación , Adulto , Androstadienos/uso terapéutico , Antiinflamatorios/uso terapéutico , Apoptosis , Regulación hacia Abajo , Eosinofilia/tratamiento farmacológico , Eosinófilos/inmunología , Eosinófilos/fisiología , Femenino , Fluticasona , Humanos , Inflamación/fisiopatología , Masculino , Metaloproteinasa 9 de la Matriz/metabolismo , Persona de Mediana Edad , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Resultado del TratamientoRESUMEN
BACKGROUND: Acetylcholine (ACh) plays an important role in smooth muscle contraction and in the development of airway narrowing; preliminary evidences led us to hypothesize that ACh might also play a role in the development of airways inflammation in chronic obstructive pulmonary disease (COPD). METHODS: We evaluated the concentrations of leukotriene B4 (LTB4) in induced sputum, and the expression of Ach M1, M2, and M3 receptors in sputum cells (SC) obtained from 16 patients with COPD, 11 smokers, and 14 control subjects. The SC were also treated with ACh and the production of LTB4 assessed in the presence or absence of a muscarinic antagonist (oxitropium). In blood monocytes, we evaluated LTB4 release and activation of the extracellular signal-regulated kinases (ERK) pathway after treatment with Ach. RESULTS: The LTB4 concentrations were higher in COPD than in controls (P < 0.01) and correlated with the number of neutrophil (P < 0.01). The M3 receptors expression was increased in COPD subjects when compared to smokers and control (P < 0.05 and 0.0001, respectively), while M2 expression resulted decreased (P < 0.05 and 0.01). The ACh-induced LTB(4) production was observed in peripheral blood monocytes, and was sensitive to ERK inhibition. Similarly, ACh significantly increased neutrophil chemotactic activity and LTB4 released from SC of COPD patients only, and these effects were blocked by pretreatment with the inhibitor of ERK pathway PD98059. CONCLUSIONS: The results obtained show that muscarinic receptors may be involved in airway inflammation in COPD subjects through ACh-induced, ERK1/2-dependent LTB4 release. Muscarinic antagonism may contribute to reduce neutrophil infiltration and activation in COPD.
Asunto(s)
Leucocitos Mononucleares/metabolismo , Leucotrieno B4/metabolismo , Neutrófilos/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/inmunología , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Receptores Muscarínicos/metabolismo , Acetilcolina/farmacología , Anciano , Proteínas de Unión al Calcio/antagonistas & inhibidores , Proteínas de Unión al Calcio/farmacología , Células Cultivadas , Quimiotaxis de Leucocito , Femenino , Flavonoides/farmacología , Humanos , Inmunohistoquímica , Leucocitos Mononucleares/efectos de los fármacos , Leucotrieno B4/análisis , Masculino , Persona de Mediana Edad , Proteína Quinasa 3 Activada por Mitógenos/efectos de los fármacos , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Proteínas Protozoarias/antagonistas & inhibidores , Proteínas Protozoarias/farmacología , Esputo/citología , Esputo/metabolismoRESUMEN
BACKGROUND: Corticosteroids play an important role in inflammation and remodelling of airways and are considered an important therapeutic target in asthma. Inflammation in asthma is characterized by a dysregulation of eosinophil apoptosis and of markers of airways remodelling. We evaluated the ability of flunisolide to inhibit in vitro the release of metalloproteinases-9 (MMP-9), tissue inhibitor metalloproteinases-1 (TIMP-1), transforming growth factor (TGF-beta) and fibronectin by sputum cells (SC) as well as to induce sputum eosinophil apoptosis. METHODS: The SC, isolated from induced sputum samples of 12 mild-to-moderate asthmatics, were cultured for 24 h in the presence or absence of flunisolide (1, 10 and 100 microM). The release of mediators was assessed by enzyme-linked immunosorbent assay (ELISA) whereas apoptosis was studied by TUNEL technique. RESULTS: Flunisolide (10 microM) significantly reduced MMP-9 and TIMP-1 (P = 0.0011 and P < 0.0001 respectively) and increased MMP-9/TIMP-1 molar ratio (P = 0.004). In addition, flunisolide decreased TGF-beta and fibronectin release by SC (P = 0.006; and P < 0.0001 respectively) and increased eosinophil apoptosis (P < 0.001). CONCLUSIONS: These results demonstrate that flunisolide may play an important role in the inhibition of airway inflammation and remodelling, by promoting the resolution of eosinophilic inflammation and by inhibiting the release of MMP-9, TIMP-1, TGF-beta and fibronectin.
Asunto(s)
Apoptosis/efectos de los fármacos , Asma/tratamiento farmacológico , Fibronectinas/metabolismo , Fluocinolona Acetonida/análogos & derivados , Fluocinolona Acetonida/farmacología , Metaloproteinasa 9 de la Matriz/metabolismo , Esputo/metabolismo , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Adulto , Asma/metabolismo , Humanos , Persona de Mediana Edad , Esputo/citología , Factor de Crecimiento Transformador beta1RESUMEN
In asthmatic subjects an imbalance between elastase and alpha1-antitrypsin (alpha1-PI) exists. This study aims to evaluate whether ageing per se affects the levels of elastase. Both young and elderly asthmatics with comparable severity and duration of disease, as well as young and elderly healthy subjects, underwent an induced sputum procedure to measure levels of elastase and alpha1-PI. The percentage of sputum neutrophils and eosinophils was higher in young and elderly asthmatics than in young and elderly controls. The levels of both total and active elastase were significantly higher in young and elderly asthmatics than in young and elderly controls, and directly correlated with the percentage of neutrophils. In addition, in both young and elderly asthmatics the levels of total and active elastase were negatively correlated with forced expiratory volume in one second values, but positively correlated with the duration of the disease. This study indicates that ageing per se does not necessarily lead to a progressive elastase/alpha1-antitrypsin imbalance in asthma, and suggests that an important variable in the development of airway remodelling in both young and elderly asthmatics is represented by the duration of the disease.
Asunto(s)
Envejecimiento/metabolismo , Asma/metabolismo , Elastasa Pancreática/metabolismo , alfa 1-Antitripsina/metabolismo , Adulto , Anciano , Envejecimiento/fisiología , Asma/patología , Asma/fisiopatología , Eosinófilos , Volumen Espiratorio Forzado , Humanos , Recuento de Leucocitos , Persona de Mediana Edad , Neutrófilos , Esputo/química , Esputo/citologíaRESUMEN
BACKGROUND: Inflammation in chronic obstructive pulmonary disease (COPD) is characterised by increased neutrophilic infiltration of the airways. Cilomilast, a novel selective phosphodiesterase 4 inhibitor in clinical development for COPD treatment, exerts anti-inflammatory effects. The ability of cilomilast to inhibit the release of neutrophil chemoattractants such as tumour necrosis factor (TNF)-alpha, interleukin (IL)-8, and granulocyte-macrophage colony stimulating factor (GM-CSF) by bronchial epithelial cells and sputum cells isolated from 10 patients with COPD, 14 normal controls, and 10 smokers was investigated. METHODS: Bronchial epithelial cells obtained by bronchial brushing and sputum cells isolated from induced sputum samples were cultured for 24 hours in the presence or absence of cilomilast (1 micro M). After incubation the supernatants were harvested and the levels of mediators measured by ELISA. Chemotactic activity in supernatants was also measured using a Boyden chamber. RESULTS: TNF-alpha and IL-8 release by bronchial epithelial cells and sputum cells was higher in patients with COPD than in controls (p<0.0001) and smokers (p<0.0001). GM-CSF was only detectable in sputum cell supernatants and its level was higher in patients with COPD than in controls and smokers (p<0.0001, respectively). Cilomilast significantly reduced TNF-alpha release by bronchial epithelial cells and sputum cells (p=0.005) and GM-CSF release by sputum cells (p=0.003), whereas IL-8 release was not statistically inhibited. Supernatants of sputum cells and bronchial epithelial cells treated with cilomilast significantly decreased neutrophil chemotaxis (p<0.006 and p<0.008, respectively). CONCLUSIONS: Cilomilast inhibits the production of some neutrophil chemoattractants by airway cells. This drug may play a role in the resolution of neutrophilic inflammation associated with COPD and cigarette smoke.
Asunto(s)
Broncodilatadores/uso terapéutico , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Interleucina-8/metabolismo , Inhibidores de Fosfodiesterasa/uso terapéutico , Enfermedad Pulmonar Obstructiva Crónica/tratamiento farmacológico , Esputo/citología , Adulto , Anciano , Ácidos Carboxílicos , Recuento de Células , Células Cultivadas , Quimiotaxis de Leucocito , Ácidos Ciclohexanocarboxílicos , Ensayo de Inmunoadsorción Enzimática , Células Epiteliales/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Nitrilos , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Mucosa Respiratoria/metabolismo , Factor de Necrosis Tumoral alfaRESUMEN
BACKGROUND: Inflammatory cells are increased in the airways of endurance athletes, but their role in causing exercise-induced respiratory symptoms and bronchoconstriction, or their possible long-term consequences, are uncertain. AIM: To put the results of athlete studies in perspective, by analysing the pathogenesis of airway cell changes and their impact on respiratory function. RESULTS: Athletes of different endurance sports at rest showed increased airway neutrophils. Elite swimmers and skiers also showed large increases in airway eosinophils and lymphocytes, possibly related to chronic, exercise-related exposure to irritants or cold and dry air, respectively. Post-exercise studies reported variable responses of airway cells to exercise, but found no evidence of inflammatory cell activation in the airways, at variance with exercise-induced neutrophil activation in peripheral blood. The increase in airway inflammatory cells in athletes can result from hyperventilation-induced increase in airway osmolarity stimulating bronchial epithelial cells to release chemotactic factors. Hyperosmolarity may also inhibit activation of inflammatory cells by causing shedding of adhesion molecules, possibly explaining why airway inflammation appears 'frustrated' in athletes. Data on exhaled nitric oxide are few and variable, not allowing conclusions about its usefulness as a marker of airway inflammation in athletes, or its role in modulating bronchial responsiveness. CONCLUSIONS: The acute and long-term effects of exercise on airway cells need further study. Airway inflammatory cells are increased but not activated in athletes, both at rest and after exercise, and airway inflammation appears to regress in athletes quitting competitions. Altogether, these findings do not clearly indicate that habitual intense exercise may be detrimental for respiratory health. Rather, airway changes may represent chronic adaptive responses to exercise hyperventilation. An improved understanding of the effects of exercise on the airways will likely have a clinical impact on sports medicine, and on the current approach to exercise-based rehabilitation in respiratory disease.
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Leucocitos/inmunología , Resistencia Física/inmunología , Mucosa Respiratoria/inmunología , Deportes , Asma Inducida por Ejercicio/inmunología , Hiperreactividad Bronquial/inmunología , Moléculas de Adhesión Celular/inmunología , Eosinófilos/citología , Humanos , Recuento de Leucocitos , Linfocitos/citología , Neutrófilos/citología , Óxido Nítrico/fisiología , Concentración OsmolarRESUMEN
Elite athletes show a high prevalence of symptoms and signs of asthma, but no study has assessed the acute effects of endurance exercise on airway cells in nonasthmatic athletes. We measured exhaled nitric oxide (NO) and collected samples of induced sputum after 3% NaCl aerosol administration for 20 min in nonasthmatic middle-aged amateur runners after the Fourth Palermo International Marathon and 6--9 wk later (habitual training period) at baseline. After the marathon, exhaled NO (n = 9 subjects) was higher [27 +/- 9 parts/billion (ppb)] than at baseline (12 +/- 4 ppb; P < 0.0005). Polymorphonuclear neutrophil (PMN) counts in induced sputum were much higher in runners (91.2 +/- 3.6% of total cells postmarathon and 78.7 +/- 9.1% at baseline) than in sedentary control subjects (9.9 +/- 5.9%; P < 0.001). Expression of L-selectin and CD11b/CD18 in sputum PMNs was lower after the race than at baseline and inversely related to the amount of exhaled NO (r = -0.66 and -0.69, respectively; P < 0.05). Our data indicate that sputum PMNs are increased in nonasthmatic runners both after a marathon and at baseline and suggest that NO may modulate exercise-associated inflammatory airway changes.
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Bronquitis/patología , Carrera , Adulto , Sangre/metabolismo , Células Sanguíneas/patología , Bronquitis/metabolismo , Bronquitis/fisiopatología , Antígenos CD18/análisis , Humanos , Selectina L/análisis , Recuento de Leucocitos , Antígeno de Macrófago-1/análisis , Masculino , Persona de Mediana Edad , Neutrófilos/patología , Óxido Nítrico , Valores de Referencia , Respiración , Pruebas de Función Respiratoria , Esputo/química , Esputo/citologíaRESUMEN
We evaluated the levels of 15(S)-hydroxyeicosatetraenoic acid [15(S)-HETE] and the expression of 15-lipoxygenase (15-LO) mRNA in induced sputum obtained from 10 control and 15 chronic bronchitis subjects. 15(S)-HETE was evaluated by reverse phase high-performance liquid chromatography separation followed by specific RIA. 15-LO mRNA expression was determined by primed in situ labeling. The levels of both soluble and cell-associated 15(S)-HETE resulted significantly higher in chronic bronchitis than in control subjects. The percentage of cells expressing 15-LO mRNA was significantly higher in chronic bronchitis than in control subjects (P < 0.01). Double staining for specific cell type markers and 15-LO mRNA showed macrophages and neutrophils positive for 15-LO, whereas similar staining of peripheral blood neutrophils did not show evidence for 15-LO expression, suggesting that expression of 15-LO in neutrophils takes place on migration into the airways. Because 15(S)-HETE inversely correlated with the percentage of neutrophils in sputum of chronic bronchitis subjects, we studied the effect of 15(S)-HETE on leukotriene B(4) (LTB(4)) production in vitro and evaluated the concentration of LTB(4) in induced sputum and the contribution of LTB(4) to the chemotactic activity of induced sputum samples ex vivo. The results obtained indicate that macrophages and neutrophils present within the airways of chronic bronchitis subjects express 15-LO mRNA; increased basal levels of 15(S)-HETE may contribute to modulate, through the inhibition of 5-lipoxygenase metabolites production, neutrophil infiltration and airway inflammation associated with chronic bronchitis.
Asunto(s)
Bronquitis/metabolismo , Ácidos Hidroxieicosatetraenoicos/metabolismo , Leucotrieno B4/biosíntesis , Enfermedades Pulmonares Obstructivas/metabolismo , Neutrófilos/metabolismo , Adulto , Anciano , Araquidonato 15-Lipooxigenasa/biosíntesis , Araquidonato 15-Lipooxigenasa/genética , Bronquitis/patología , Recuento de Células , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Quimiotaxis de Leucocito/efectos de los fármacos , Enfermedad Crónica , Humanos , Ácidos Hidroxieicosatetraenoicos/análisis , Ácidos Hidroxieicosatetraenoicos/farmacología , Inmunohistoquímica , Hibridación in Situ , Ionóforos/farmacología , Antagonistas de Leucotrieno/farmacología , Enfermedades Pulmonares Obstructivas/patología , Macrófagos/metabolismo , Persona de Mediana Edad , Neutrófilos/efectos de los fármacos , Neutrófilos/patología , ARN Mensajero/biosíntesis , Esputo/química , Esputo/citología , Esputo/metabolismoRESUMEN
BACKGROUND: Recent evidence shows that 15(S)-hydroxy-eicoisatetraenoic acid (15[S]-HETE) can be released and rapidly reincorporated into cellular lipids. These mechanisms exert several immunoregulatory functions that may be relevant in airway inflammation. OBJECTIVE: Our purpose was to evaluate the levels of both soluble and cell-associated 15(S)-HETE and to examine 15-lipoxygenase (15-LO) messenger RNA (mRNA) expression in sputum samples obtained from 10 control and 18 asthmatic subjects. METHODS: Levels of 15(S)-HETE were measured by reverse-phase HPLC separation followed by RIA in supernatants and in cell membrane-extracted phospholipids after acid hydrolysis. 15-LO mRNA was evaluated by primed in situ hybridization (PRINS). Combined immunocytochemistry and PRINS was used to identify the phenotype of cells bearing 15-LO transcripts. RESULTS: Levels of both soluble and cell-associated 15(S)-HETE were higher in asthmatic than in control subjects (P <.0001). The percentage of cells expressing 15-LO mRNA was higher in asthmatic than in control subjects (P <.01). On double staining for specific cell-type markers and 15-LO mRNA, macrophages were the major source for 15-LO. CONCLUSION: This study shows that the induced sputum technique allows the evaluation of 15-LO activity and that soluble, cell-associated 15(S)-HETE and 15-LO levels are higher in asthmatic than in control subjects. In addition, this study indicates that, in induced sputum, airway macrophages are the major source of 15(S)-HETE in asthma.
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Araquidonato 15-Lipooxigenasa/genética , Asma/metabolismo , Ácidos Hidroxieicosatetraenoicos/metabolismo , Adulto , Anciano , Recuento de Células , Volumen Espiratorio Forzado , Humanos , Inmunohistoquímica , Hibridación in Situ , Persona de Mediana Edad , ARN Mensajero/metabolismo , Saliva/citología , Solubilidad , Esputo/química , Esputo/citología , Esputo/enzimologíaRESUMEN
Asthma and chronic bronchitis are inflammatory diseases with extracellular matrix (ECM) remodeling and collagen deposition. Collagen homeostasis is controlled by metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). We evaluated MMP and TIMP balance in induced sputum of 10 control, 31 untreated asthmatic, and 16 chronic bronchitic subjects. We first performed zymographic analysis to identify the profile of MMPs. Zymography revealed a similar MMPs profile in all populations studied and that MMP-9 was the major enzyme released. We then measured, using enzyme immunoassay, the concentrations of MMP-9 and of its inhibitor TIMP-1 and evaluated whether airflow limitation may be associated with an imbalance between these enzymes. MMP-9 and TIMP-1 concentrations were greater in sputum of patients with asthma and chronic bronchitis than in control subjects. The molar ratio between MMP-9 and TIMP-1 was lower in asthmatics and chronic bronchitics than in control subjects, and positively correlated with FEV1 values. In asthma, MMP-9 levels were significantly correlated with the number of macrophages and neutrophils. This study shows that airway inflammation in asthma and chronic bronchitis is associated with an imbalance between MMP-9 and TIMP-1 which may have a role in the pathogenesis of ECM remodeling and airflow obstruction.
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Obstrucción de las Vías Aéreas/metabolismo , Asma/metabolismo , Bronquitis/metabolismo , Colagenasas/análisis , Inhibidores de Proteasas/análisis , Esputo/química , Inhibidor Tisular de Metaloproteinasa-1/análisis , Adolescente , Adulto , Anciano , Obstrucción de las Vías Aéreas/enzimología , Obstrucción de las Vías Aéreas/patología , Obstrucción de las Vías Aéreas/fisiopatología , Asma/enzimología , Asma/patología , Asma/fisiopatología , Bronquitis/enzimología , Bronquitis/patología , Bronquitis/fisiopatología , Recuento de Células , Enfermedad Crónica , Colágeno/metabolismo , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Matriz Extracelular/metabolismo , Volumen Espiratorio Forzado/fisiología , Homeostasis/fisiología , Humanos , Recuento de Leucocitos , Macrófagos/patología , Metaloproteinasa 9 de la Matriz , Persona de Mediana Edad , Neutrófilos/patología , Ventilación Pulmonar/fisiología , Dodecil Sulfato de Sodio , Esputo/citología , Esputo/enzimología , TensoactivosRESUMEN
Asthma and chronic bronchitis are inflammatory diseases associated with remodeling of the extracellular matrix (ECM). Elastin, a major component of the ECM in the airways, has been previously found to be disrupted in asthma and chronic bronchitis. This study was aimed at evaluating whether elastin disruption might be associated with an imbalance between elastase (active and total) and alpha1-proteinase inhibitor (alpha1-PI), the main inhibitor of elastase. We measured elastase and alpha1-PI in induced sputum obtained from 16 control subjects, 10 healthy smokers, 19 asthmatic patients, and 10 chronic bronchitis patients. We also assessed the possible origin of elastase, evaluating its levels in sputum with reference to differential cell counts. We found that in induced sputum obtained from asthmatic and chronic bronchitis patients, the levels of both total and active elastase were significantly increased as compared with those of control subjects and healthy smokers and were significantly correlated with the percentage of neutrophils. In addition, in asthma and chronic bronchitis patients, the levels of active and total elastase were inversely correlated with the degree of airway obstruction as assessed from FEV1 values. This study shows that airway inflammation in asthma and chronic bronchitis is associated with high levels of active elastase, which may play a role in the pathogenesis of airway remodeling.
Asunto(s)
Asma/metabolismo , Elastasa Pancreática/metabolismo , Esputo/metabolismo , alfa 1-Antitripsina/metabolismo , Adulto , Anciano , Asma/patología , Asma/fisiopatología , Bronquitis/metabolismo , Bronquitis/fisiopatología , Recuento de Células , Enfermedad Crónica , Volumen Espiratorio Forzado/fisiología , Humanos , Persona de Mediana Edad , Valores de Referencia , Saliva/citología , Albúmina Sérica/metabolismo , Fumar , Esputo/citologíaRESUMEN
To evaluate the release of catecholamines and their relationship with systemic blood pressure (BP) in normotensive patients with obstructive sleep apnea syndrome (OSAS), diurnal and nocturnal urinary norepinephrine (NE) and epinephrine (E) excretion in 12 normal subjects and in 10 OSAS patients were compared; in addition, nocturnal NE and E excretion were measured in the patients while receiving short-term CPAP. Blood pressure was continuously monitored in the patients during both nights of urine collection. In normal subjects, both NE and E excretion decreased from day to night. In the patients without CPAP, only NE excretion decreased at night, and BP increased from wakefulness to sleep; both NE and E excretion were higher in patients than in normal subjects. With CPAP, which prevented apneas, only E excretion decreased with respect to the previous night, while BP no longer increased during sleep. The extent of nocturnal E decrease with CPAP was not correlated to BP variations. These results suggest that in normotensive OSAS subjects, sympathetic nervous system activity, based on NE excretion, is continuously increased and is not affected by short-term CPAP treatment. Conversely, adrenal activity, based on E excretion, is also increased, but it tends to be normalized by short-term CPAP. No clear relationship could be found between sympatho-adrenal behavior and BP during sleep.
Asunto(s)
Presión Sanguínea , Catecolaminas/orina , Síndromes de la Apnea del Sueño/orina , Adulto , Análisis de Varianza , Ritmo Circadiano , Diástole , Epinefrina/orina , Femenino , Humanos , Masculino , Persona de Mediana Edad , Norepinefrina/orina , Respiración con Presión Positiva , Síndromes de la Apnea del Sueño/epidemiología , Síndromes de la Apnea del Sueño/fisiopatología , Síndromes de la Apnea del Sueño/terapia , SístoleRESUMEN
Immunoglobulin (Ig) levels increase in the lower respiratory tract of patients with pulmonary sarcoidosis. We evaluated the effects of prednisone therapy upon Ig concentrations in bronchoalveolar lavage (BAL) fluid of ten patients with active disease (greater than 30% T-lymphocytes in BAL and positive 67Gallium (67Ga) lung scan). Therapy significantly lowered T-lymphocyte percentages in BAL and 67Ga lung scan indices and was followed by a slight improvement of the studied functional parameters. Biochemical analysis of BAL showed a significant decrease of both IgG/albumin (baseline 1.24 +/- 0.21; after therapy 0.40 +/- 0.12) and IgA/albumin (baseline 0.55 +/- 0.07; after therapy 0.14 +/- 0.03) ratios in all patients. Conversely, comparisons of IgM/albumin ratios did not show any change over the study period (baseline 0.05 +/- 0.01; after therapy 0.06 +/- 0.03). Thus oral steroid treatment suppresses the alveolitis of pulmonary sarcoidosis, as shown not only by the reduction of lung T-cells and 67Ga lung uptake, but also by the decreased Ig levels in the alveolar spaces.
Asunto(s)
Líquido del Lavado Bronquioalveolar/análisis , Inmunoglobulinas/análisis , Enfermedades Pulmonares/tratamiento farmacológico , Prednisona/uso terapéutico , Sarcoidosis/tratamiento farmacológico , Adulto , Líquido del Lavado Bronquioalveolar/citología , Femenino , Humanos , Masculino , Linfocitos T/clasificaciónRESUMEN
Ascitic fluid pH and arterial-ascitic fluid pH gradient were compared to ascitic fluid polymorphonuclear cell count in 84 patients with cirrhotic ascites and in 12 with malignant ascites to assess their role as diagnostic tests for spontaneous bacterial peritonitis and to clarify the relationship between ascitic fluid pH and lactate. Ascitic fluid pH was significantly lower (pH 7.30) in spontaneous bacterial peritonitis (n = 18) and probable spontaneous bacterial peritonitis (n = 12) than in sterile ascites (pH 7.41; n = 54). Since blood pH levels were not different in the presence of infection, arterial-ascitic fluid pH gradient was significantly higher in spontaneous bacterial peritonitis and probable spontaneous bacterial peritonitis than in sterile ascites (0.12 vs. 0.02). The close correlations between arterial-ascitic pH gradient and lactate (r = 0.77, p less than 0.0001), lactate and bicarbonate gradient (r = 0.64, p = 0.003) and arterial-ascitic pH gradient and pCO2 gradient (r = -0.90, p less than 0.0001) suggest that the low ascitic fluid pH may be due to an increase in lactate and CO2. Patients with Escherichia coli-induced spontaneous bacterial peritonitis had significantly lower ascitic fluid pH and higher lactate than those with spontaneous bacterial peritonitis by other organisms. Values of ascitic fluid pH, lactate and arterial-ascitic fluid pH gradient in malignant ascites were similar to those of spontaneous bacterial peritonitis and probable spontaneous bacterial peritonitis. Cutoff points, selected by receiver operating characteristic curves analysis, of 450 per mm3 for polymorphonuclear cells and of 0.07 for arterial-ascitic fluid pH gradient, allow high positive and negative predictive values for spontaneous bacterial peritonitis.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Líquido Ascítico/metabolismo , Infecciones Bacterianas/diagnóstico , Lactatos/análisis , Peritonitis/diagnóstico , Anciano , Ascitis/etiología , Líquido Ascítico/patología , Infecciones Bacterianas/complicaciones , Infecciones por Escherichia coli/complicaciones , Infecciones por Escherichia coli/diagnóstico , Femenino , Humanos , Concentración de Iones de Hidrógeno , Ácido Láctico , Recuento de Leucocitos , Cirrosis Hepática/complicaciones , Masculino , Persona de Mediana Edad , Neutrófilos , Neoplasias Peritoneales/complicaciones , Peritonitis/complicacionesRESUMEN
Cellular and biochemical analyses of bronchoalveolar lavage (BAL) were performed in 8 normal subjects and in 18 patients with pulmonary sarcoidosis. The patients were divided into two groups, according to the intensity of the alveolitis as assessed by lung T-lymphocyte percentage and by 67Ga lung scan. High-intensity alveolitis (HIA) patients had an increased ratio of OKT4-positive: OKT8-positive T cells in their lungs, but not in their blood, compared to low-intensity alveolitis (LIA) patients and to controls. Biochemical analyses of BAL showed that HIA patients had increased albumin and IgG concentrations compared to LIA patients and to controls. IgA concentrations were more elevated in sarcoid patients than in controls, with no difference between the two groups of patients. No differences were detected in IgM concentrations between the three groups of subjects. The levels of different Ig classes were then calculated as a ratio with respect to albumin in order to determine whether their presence in BAL fluid was due to increased alveolar-capillary 'leak'. The IgG:albumin ratio was significantly higher in HIA patients compared to LIA patients and to controls, whereas comparison of the IGA: albumin and IgM: albumin ratios showed no significant differences between the three groups of subjects. These findings suggest that alveolar-capillary permeability is increased in pulmonary sarcoidosis and provide evidence that local IgG production is enhanced in active states of this disease.