RESUMEN
Pseudomonas aeruginosa (PA) airway infection and bronchial blood vessel proliferation are features of bronchiectasis. Because vascular endothelial growth factor (VEGF)-A regulates angiogenesis, we hypothesised that PA infection induces VEGF synthesis in epithelium and peribronchial angiogenesis. Because epidermal growth factor receptor (EGFR) activation regulates VEGF synthesis in cancer, we also evaluated the roles of EGFR. Airway epithelial cells were incubated for 24 h with PA supernatants and VEGF concentrations were measured in culture medium by ELISA. C57BL/6N mice were instilled intratracheally with sterile agarose beads or with agarose beads coated with the PA strain PAO1 (mean ± sem 6 × 10(5) ± 3 × 10(5) cfu · animal(-1)), with or without the EGFR inhibitor AG1478 (12.5 mg · kg(-1) · day(-1) intraperitoneally). Epithelial immunostaining for VEGF and phosphorylated EGFR, and peribronchial vascularity, were quantified using morphometric analysis. VEGF expression was further assessed by western blot in mouse lung homogenates. PA supernatants induced dose-dependent VEGF synthesis in cultured airway epithelial cells, effects which were prevented by EGFR antagonists. In mice, persistent PAO1 infection increased immunostaining for VEGF and phosphorylated EGFR in airway epithelium, and resulted in increased peribronchial vascularity within 7 days. These effects were reduced by EGFR inhibition. Persistent PA infection induced VEGF synthesis in airway epithelium and peribronchial angiogenesis, at least in part via EGFR-dependent mechanisms.
Asunto(s)
Infecciones por Pseudomonas/metabolismo , Pseudomonas aeruginosa/metabolismo , Mucosa Respiratoria/metabolismo , Mucosa Respiratoria/microbiología , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Animales , Bronquiectasia/metabolismo , Bronquiectasia/microbiología , Carcinoma Mucoepidermoide , Línea Celular Tumoral , Modelos Animales de Enfermedad , Receptores ErbB/metabolismo , Femenino , Humanos , Técnicas In Vitro , Neoplasias Pulmonares , Ratones , Ratones Endogámicos C57BL , Neovascularización Patológica/metabolismo , Neovascularización Patológica/microbiología , Circulación Pulmonar/fisiología , Mucosa Respiratoria/irrigación sanguínea , Transducción de Señal/fisiología , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
We examined the effects of acute exposure to cigarette smoke on the airway responses to substance P in anesthetized guinea pigs and on the activity of airway neutral endopeptidase (NEP). After exposure to air or to cigarette smoke we measured the change in total pulmonary resistance (RL) induced by increasing concentrations of aerosolized substance P in the absence or presence of the NEP inhibitor phosphoramidon. In the absence of phosphramidon the bronchoconstrictor responses to substance P were greater in cigarette smoke-exposed guinea pigs than in air-exposed animals. Phosphoramidon did not further potentiate the responses to substance P in smoke-exposed guinea pigs, whereas it did so in air-exposed animals. In the presence of phosphoramidon, bronchoconstrictor responses to substance P in animals exposed to air or to cigarette smoke were not different. Aerosols of SOD delivered before cigarette smoke exposures dramatically reduced smoke-induced hyperresponsiveness to substance P, whereas heat-inactivated SOD had no effect on smoke-induced hyper-responsiveness to substance P. Cigarette smoke solution inhibited NEP activity from tracheal homogenate in a concentration-dependent fashion, an inhibitory effect that was mostly due to the gas phase of the smoke, but not to nicotine. The mild chemical oxidant N-chlorosuccinimide mimicked the concentration-dependent inhibitory effect of smoke solution on airway NEP activity. We conclude that cigarette smoke causes enhanced airway responsiveness to substance P in vivo by inactivating airway NEP. We suggest that cigarette smoke-induced inhibition of airway NEP is due to effects of free radicals.
Asunto(s)
Resistencia de las Vías Respiratorias , Bronquios/enzimología , Neprilisina/metabolismo , Fumar/efectos adversos , Sustancia P/administración & dosificación , Resistencia de las Vías Respiratorias/efectos de los fármacos , Animales , Pruebas de Provocación Bronquial , Capsaicina/administración & dosificación , Activación Enzimática , Radicales Libres , Glicopéptidos/administración & dosificación , Cobayas , Masculino , Nicotina/administración & dosificación , Soluciones , Succinimidas/administración & dosificación , Superóxido Dismutasa/administración & dosificaciónRESUMEN
We studied the perception of bronchoconstriction in asthmatic subjects who were randomly treated with inhaled beta 2 agonist given either alone (n = 9) or associated with inhaled corticosteroids (n = 9). Methacholine and bradykinin challenges, bronchoalveolar lavage, and bronchial biopsies were performed in all subjects. After each dose of agonist, breathlessness was assessed using a visual analog scale (VAS) and the forced expiratory volume in 1 s (FEV1) was measured. The relationship between VAS scores and FEV1 and the slope of the regression line of VAS scores on the corresponding FEV1 (VAS/FEV1 slope) were analyzed for each agonist. Subjects without corticosteroids had good perception of methacholine but poor perception of bradykinin-induced bronchoconstriction. In subjects with corticosteroids, bronchoconstriction was well perceived whatever the agonist. VAS/FEV1 slopes for bradykinin but not for methacholine correlated negatively with the magnitude of eosinophilic inflammation in airway mucosa. VAS/FEV1 slopes for each agonist correlated positively with the percentage of basement membrane covered by airway epithelium. We conclude that in asthmatic patients perception of bronchoconstriction is related to eosinophilic inflammation and to epithelial damage in airways and that corticosteroid treatment is associated with improved perception of bronchoconstriction induced by bradykinin, a mediator endogenously produced in asthma.
Asunto(s)
Corticoesteroides/uso terapéutico , Asma/fisiopatología , Bronquios/patología , Broncoconstricción/efectos de los fármacos , Eosinófilos/patología , Inflamación/fisiopatología , Adulto , Anciano , Asma/tratamiento farmacológico , Bradiquinina/farmacología , Epitelio/patología , Femenino , Humanos , Inflamación/patología , Masculino , Cloruro de Metacolina/farmacología , Persona de Mediana Edad , PercepciónRESUMEN
Nitric oxide (NO) plays a central role in many airway physiological functions, and its production appears to be related with progression of lung disease in patients with cystic fibrosis (CF). However, underlying mechanisms which specifically link NO and CF-related lung disease remain unclear. Following in vitro and animal studies suggesting a role for NO in ion transport in various epithelia, this work investigates the relationship between transepithelial baseline potential difference (BPD), an index of airway ion transport, and exhaled NO in the airways of adult patients with CF. Association with other phenotypic traits, lung function tests and CFTR genotype was also assessed. Using simple linear regression, F(E)NO and transepithelial BPD values were significantly inversely correlated (p<0.001, r=-0.53). Polynomial analysis evidenced an asymptotic relationship between F(E)NO and BPD values, yielding a plateau for absolute BPD values above 50 mV. This relation was not altered by adjustment for clinical and genetic characteristics of the patients. The relationship between exhaled NO and transepithelial BPD suggests that low NO concentrations likely worsens airway ion transport impairment resulting from CFTR defect. These results fit with experimental studies that suggest the inhibitory effect of NO on sodium absorption, which is the main determinant of airway basal transepithelial conductance.
Asunto(s)
Fibrosis Quística/metabolismo , Fibrosis Quística/fisiopatología , Mucosa Nasal/fisiopatología , Óxido Nítrico/metabolismo , Adulto , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Femenino , Genotipo , Humanos , Modelos Lineales , Masculino , Potenciales de la Membrana/fisiología , Persona de Mediana Edad , Pruebas de Función RespiratoriaRESUMEN
To determine whether neutral endopeptidase (NEP), also called enkephalinase (EC 3.4.24.11), modulates the effects of exogenous and endogenous tachykinins in vivo, we studied the effects of aerosolized phosphoramidon, a specific NEP inhibitor, on the responses to aerosolized substance P (SP) and on the atropine-resistant response to vagus nerve stimulation (10 V, 5 ms for 20 s) in guinea pigs. SP alone (10(-7) to 10(-4) M; each concentration, 7 breaths) caused no change in total pulmonary resistance (RL, P greater than 0.5). Phosphoramidon (10(-4) M, 90 breaths) caused no change either in base-line RL (P greater than 0.5) or in the response to aerosolized acetylcholine (P greater than 0.5). However, in the presence of phosphoramidon, SP (7 breaths) produced a concentration-dependent increase in RL at concentrations greater than or equal to 10(-5) M (P less than 0.001). Phosphoramidon (10(-7) to 10(-4) M; each concentration, 90 breaths) induced a concentration-dependent potentiation of SP-induced bronchoconstriction (10(-4) M, 7 breaths; P less than 0.01). Vagus nerve stimulation (0.5-3 Hz), in the presence of atropine, induced a frequency-dependent increase in RL (P less than 0.001). Phosphoramidon potentiated the atropine-resistant responses to vagus nerve stimulation (P less than 0.001) at frequencies greater than 0.5 Hz. The tachykinin antagonist [D-Arg1,D-Pro2,D-Trp7,9,Leu11]-substance P abolished the effects of phosphoramidon on the atropine-resistant response to vagus nerve stimulation (2 Hz, P less than 0.005). NEP-like activity in tracheal homogenates of guinea pig was inhibited by phosphoramidon with a concentration producing 50% inhibition of 5.3 +/- 0.8 nM.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Bronquios/fisiología , Neprilisina/fisiología , Resistencia de las Vías Respiratorias/efectos de los fármacos , Animales , Atropina/farmacología , Bronquios/efectos de los fármacos , Estimulación Eléctrica , Glicopéptidos/farmacología , Cobayas , Masculino , Neprilisina/antagonistas & inhibidores , Sustancia P/farmacología , Taquicininas/farmacología , Nervio Vago/fisiologíaRESUMEN
To determine the role of endogenous neutral endopeptidase (NEP) (also called enkephalinase, EC 3.4.24.11) in regulating neurotensin-induced airway contraction, we used phosphoramidon, a specific NEP inhibitor, in the guinea pig. In studies in vitro, neurotensin and the COOH-terminal fragment neurotensin-(8-13) contracted strips of bronchial smooth muscle in a concentration-dependent fashion (P less than 0.001). In contrast, the NH2-terminal fragment neurotensin-(1-11) and the COOH-terminal fragment neurotensin-(12-13), the main fragments of neurotensin hydrolysis by NEP, had no effect. Phosphoramidon (10(-5) M) did not change resting tension but shifted the concentration-response curves to neurotensin to lower concentrations (P less than 0.001), whereas inhibitors of kininase II, aminopeptidases, serine proteases, and carboxypeptidase N were without effect. Removing the epithelium increased the contractile response to neurotensin (P less than 0.001), and phosphoramidon further increased the response to neurotensin in these tissues (P less than 0.001). Similar results were obtained in studies in vivo using aerosolized neurotensin and phosphoramidon. These results suggest that endogenous NEP in the airways modulates the effects of neurotensin on airway smooth muscle contraction by inactivating the peptide.
Asunto(s)
Resistencia de las Vías Respiratorias/efectos de los fármacos , Bronquios/fisiología , Contracción Muscular/efectos de los fármacos , Neprilisina/metabolismo , Neurotensina/farmacología , Aerosoles , Animales , Bronquios/efectos de los fármacos , Epitelio/fisiología , Cobayas , Técnicas In Vitro , Masculino , Músculo Liso/efectos de los fármacos , Músculo Liso/fisiología , Neurotensina/administración & dosificación , Valores de ReferenciaRESUMEN
We examined the effects of viral respiratory infection by Sendai virus on airway responsiveness to tachykinins in guinea pigs. We measured the change in total pulmonary resistance induced by substance P or capsaicin in the presence or absence of the neutral endopeptidase inhibitor, phosphoramidon, in infected and in noninfected animals. In the absence of phosphoramidon, the bronchoconstrictor responses to substance P and to capsaicin were greater in infected than in noninfected animals. Phosphoramidon did not further potentiate the responses to substance P and to capsaicin in the infected animals, whereas it did so in noninfected animals. Studies performed in vitro showed that nonadrenergic noncholinergic bronchial smooth muscle responses to electrical field stimulation were also increased in tissues from infected animals and that phosphoramidon increased the response of tissues from noninfected animals greatly but increased the responses of tissues from infected animals only slightly. Responses to acetylcholine were unaffected by viral infection. Neutral endopeptidase activity was decreased by 40% in the tracheal epithelial layer of the infected animals. We suggest that respiratory infection by Sendai virus causes enhanced airway responsiveness to tachykinins by decreasing neutral endopeptidase-like activity in the airway epithelium.
Asunto(s)
Bronquios/efectos de los fármacos , Neprilisina/fisiología , Infecciones por Paramyxoviridae/fisiopatología , Infecciones del Sistema Respiratorio/fisiopatología , Taquicininas/farmacología , Animales , Bronquios/fisiopatología , Capsaicina/farmacología , Constricción Patológica , Estimulación Eléctrica , Glicopéptidos/farmacología , Cobayas , Masculino , Músculo Liso/fisiopatología , Neprilisina/antagonistas & inhibidores , Virus de la Parainfluenza 1 Humana , Infecciones del Sistema Respiratorio/etiología , Sustancia P/farmacologíaRESUMEN
We studied 10 healthy nonsmokers and 8 healthy smokers, in both the upright and supine position, to investigate whether regional differences in respiratory clearance of technetium-99m-labeled diethylenetriamine pentaacetic acid 99mTc-DTPA (RC-DTPA) existed and to assess the influence of posture and smoking on the regional RC-DTPA. RC-DTPA was assessed by the lung clearance rates (%/min) of aerosolized 99mTc-DTPA (0.8 micron MMD; 2.4 GSD), using data corrected for recirculating radioactivity, in the upper (zone 1), middle (zone 2), and lower (zone 3) posterior lung fields. In nonsmokers, RC-DTPA in zone 1 was faster than in zone 2 or 3 in both the upright (P less than 0.001) and supine positions (P less than 0.0). No effect was produced by changes in posture on the regional RC-DTPA. In smokers, RC-DTPA was increased in all zones compared with the nonsmokers (P = 0.004), with a further increase in RC-DTP in zone 1 in the upright posture compared with the other regions (P less than 0.001). We conclude that in nonsmokers regional RC-DTPA is faster in zone 1 than in other zones, and this is not related to recirculation of radioactivity; posture does not modify the regional RC-DTPA of nonsmokers; smoking increases RC-DTPA in all zones and more in zone 1 in the upright posture.
Asunto(s)
Ácido Pentético/metabolismo , Postura , Sistema Respiratorio/metabolismo , Fumar , Adulto , Aerosoles , Femenino , Humanos , Masculino , Tecnecio , Distribución TisularRESUMEN
Oxidants have been implicated in the pathogenesis of many inflammatory airway diseases. Neutral endopeptidase (also called enkephalinase, EC 3.4.24.11) is a peptidase that is involved in the degradation of several proinflammatory peptides, such as tachykinins and kinins. Indirect evidence suggests that airway neutral endopeptidase is inactivated by oxidants. To determine whether hydrogen peroxide inactivates neutral endopeptidase, we studied the activity of this peptidase in washed crude preparations of membranes from guinea pig lungs. Washed crude membrane preparations were exposed to increasing concentrations of hydrogen peroxide (1.25-25 mM) in the presence or absence of two different concentrations of catalase (300 and 700 U/mL). Neutral endopeptidase activity was inhibited by hydrogen peroxide in a concentration-dependent fashion (p = .0001). Addition of catalase prevented, in a concentration-dependent fashion, the inhibition of neutral endopeptidase induced by hydrogen peroxide (p = .0001). Mannitol (40 mM) and L-methionine (20 mM) did not prevent inhibition of neutral endopeptidase induced by hydrogen peroxide (2.5 mM). It can be concluded that neutral endopeptidase is inactivated by hydrogen peroxide, an effect that is prevented by catalase. Hydrogen peroxide-induced inactivation of neutral endopeptidase is not mediated by spontaneous generation of either hydroxyl radical or hypochlorous acid in the membrane preparation. Our results suggest that neutral endopeptidase inactivation may occur in airway diseases associated with exposure to or production of oxidants.
Asunto(s)
Peróxido de Hidrógeno/farmacología , Pulmón/enzimología , Neprilisina/antagonistas & inhibidores , Animales , Catalasa/farmacología , Inducción Enzimática , Cobayas , Radical Hidroxilo/metabolismo , Ácido Hipocloroso/metabolismo , Masculino , Neprilisina/metabolismoRESUMEN
We studied the effects of aerosolized DL-2-mercaptomethyl-3-guanidino-ethylthiopropanoic acid (MGTA) (10(-4) M, 90 breaths), a specific inhibitor of carboxypeptidase B-type enzymes, on changes in total pulmonary resistance (RL) induced by aerosolized capsaicin (10(-7) to 10(-4) M; 10 breaths at each concentration) and vagus nerve stimulation (5 V, 5 ms, for 20 s at frequencies varying from 2 to 10 Hz) in anesthetized, atropinized, and ventilated guinea pigs. We also studied the effect of aerosolized MGTA on the bronchoconstrictor response to either aerosolized substance P, neurokinin A (10(-7) to 10(-4) M; 10 breaths at each concentration), and carbachol (10(-5) to 2 x 10(-4) M; 10 breaths at each concentration) or to i.v. administration of neurokinin A (10(-11) to 10(-8) mol/kg), bradykinin (10(-10) to 10(-7) mol/kg), and histamine (10(-8) to 10(-6) mol/kg). Although aerosolized MGTA caused no change in basal RL (P > 0.5), it did potentiate the noncholinergic bronchoconstrictor response to capsaicin (n = 5; P < 0.001) as well as to vagus nerve stimulation (n = 5; P = 0.001). In contrast, MGTA did not potentiate the bronchoconstrictor response to either aerosolized substance P, neurokinin A, and carbachol or to i.v. administration of neurokinin A, histamine, and bradykinin. Carboxypeptidase activity cleaving C-terminal arginine or lysine was found in the membrane preparations of trachea and lung from guinea pigs. The membrane-bound carboxypeptidase activity was maximal at pH 7.0 and was enhanced by the presence of CoCl2 (1 mM) in both the tracheal and lung tissue.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Broncoconstricción , Metaloendopeptidasas/metabolismo , Ácido 3-Mercaptopropiónico/análogos & derivados , Ácido 3-Mercaptopropiónico/farmacología , Animales , Bradiquinina/farmacología , Capsaicina/farmacología , Estimulación Eléctrica , Proteínas Ligadas a GPI , Cobayas , Histamina/farmacología , Masculino , Metaloendopeptidasas/antagonistas & inhibidores , Neuroquinina A/farmacología , Receptores Colinérgicos/metabolismo , Sustancia P/farmacología , Nervio Vago/fisiologíaRESUMEN
We investigated the effects of tachykinins on the respiratory clearance of 99mTc-DTPA (RC-DTPA) in anesthetized and ventilated guinea pigs. We measured the change in RC-DTPA and in maximal pulmonary insufflation pressure (PIPmax) induced by substance P, neurokinin A, and capsaicin. Substance P, neurokinin A, and capsaicin increased both PIPmax and RC-DTPA in a concentration-dependent fashion. Substance-P- and capsaicin-induced increases in RC-DTPA were unaffected by pretreatment with atropine. Bilateral vagotomy attenuated substance-P-induced change in both RC-DTPA and PIPmax by approximately 70 and 50%, respectively. Capsaicin-induced change in RC-DTPA and PIPmax were slightly but not significantly reduced by bilateral vagotomy. The bronchodilator, salbutamol, dramatically reduced increase in RC-DTPA and in PIPmax induced by substance P, neurokinin A, and capsaicin, but it had no effect on increases in RC-DTPA and PIPmax generated by application of a positive end-expiratory pressure. We conclude that (1) tachykinins increase respiratory clearance to the solute, and (2) tachykinin-induced increase in RC-DTPA is not mediated by cholinergic neurotransmission but rather by the bronchoconstrictor effect of neuropeptides.
Asunto(s)
Capsaicina/farmacología , Pulmón/diagnóstico por imagen , Taquicininas/farmacología , Pentetato de Tecnecio Tc 99m , Albuterol/farmacología , Animales , Atropina/farmacología , Broncoconstricción/efectos de los fármacos , Permeabilidad Capilar/fisiología , Relación Dosis-Respuesta a Droga , Cobayas , Pulmón/fisiología , Masculino , Respiración con Presión Positiva , Alveolos Pulmonares/metabolismo , Cintigrafía , VagotomíaRESUMEN
We studied the effect of nasal administration of capsaicin in eight patients with allergic rhinitis and eight healthy subjects. We also studied the effect of colchicine, a drug known to inhibit microtubular axonal transport of peptides, on nasal response to capsaicin in these subjects. Colchicine or placebo was administered orally in a double-blind, randomized, crossover manner with a 35 day wash-out interval. Nasal challenge was performed on the last day of each period of treatment, using increasing doses of capsaicin (10(-9) to 3 x 10(-5) mmol). Capsaicin induced a dose-dependent decrease in nasal airflow conductance (active posterior rhinomanometry) (p < 0.002) that was greater in patients with allergic rhinitis (0.40 +/- 0.02 to 0.20 +/- 0.03) than in healthy subjects (0.44 +/- 0.01 to 0.35 +/- 0.02) (p = 0.0001). Capsaicin provoked a greater number of sneezes in patients with allergic rhinitis than in healthy subjects (p < 0.001), but the amount of nasal secretions was similar in these two groups of subjects. In nasal lavage fluid, capsaicin induced an increase in total, epithelial, and neutrophil cell counts in patients with allergic rhinitis (each comparison, p < 0.05), but not in healthy subjects. Capsaicin induced a slight, although not significant, increase in the concentration of elastase in nasal lavage fluid in patients with allergic rhinitis (p = 0.07), but not in healthy subjects. Albumin concentration decreased in nasal lavage fluid in both groups of subjects (p < 0.05). The tendency of capsaicin to increase neutrophil elastase in nasal lavage fluid of patients with allergic rhinitis was not observed after treatment with colchicine.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Capsaicina/farmacología , Colchicina/farmacología , Mucosa Nasal/efectos de los fármacos , Pruebas de Provocación Nasal , Rinitis Alérgica Perenne/fisiopatología , Adulto , Albúminas/análisis , Recuento de Células , Estudios Cruzados , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Humanos , Elastasa de Leucocito/análisis , Masculino , Líquido del Lavado Nasal/química , Activación Neutrófila/efectos de los fármacos , Neutrófilos/enzimología , Elastasa Pancreática/análisisRESUMEN
We studied the role of neutral endopeptidase (NEP) and kininase II (angiotensin-converting enzyme; ACE) in the modulation of exogenous substance P (SP)-induced nasal response in normal subjects and in patients with allergic rhinitis. We measured the nasal conductance in response to increasing doses of SP 2 h after oral administration of either placebo or the ACE inhibitor, cilazapril (5 mg), or the NEP inhibitor, acetorphan (300 mg), given in a randomized, double-blind, cross-over manner. We performed three separate studies: acetorphan versus placebo and cilazapril versus placebo, in normal subjects (n = 6 and n = 8, respectively), and acetorphan versus cilazapril versus placebo in patients with allergic rhinitis (n = 6). In normal as well as in rhinitic subjects, SP decreased nasal conductance in a dose-dependent fashion (p < 0.001). With placebo, the decrease in nasal conductance in normal subjects was similar to that in patients with allergic rhinitis (p > 0.5). In normal subjects, acetorphan potentiated the decrease in nasal conductance (p < 0.001), whereas cilazapril did not (p = 0.12). In patients with allergic rhinitis, the decrease in nasal conductance was potentiated by acetorphan (p < 0.001) and by cilazapril (p < 0.001). With acetorphan, the decrease in nasal conductance was not different in patients with allergic rhinitis and in normal subjects (p > 0.9). Conversely, with cilazapril, the nasal response to SP was greater in patients with allergic rhinitis than in normal subjects (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Cilazapril/uso terapéutico , Obstrucción Nasal/tratamiento farmacológico , Obstrucción Nasal/fisiopatología , Neprilisina/antagonistas & inhibidores , Neprilisina/fisiología , Peptidil-Dipeptidasa A/fisiología , Rinitis Alérgica Perenne/complicaciones , Tiorfan/análogos & derivados , Administración Intranasal , Administración Oral , Adulto , Resistencia de las Vías Respiratorias/efectos de los fármacos , Análisis de Varianza , Cilazapril/farmacología , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Sinergismo Farmacológico , Humanos , Masculino , Obstrucción Nasal/inducido químicamente , Obstrucción Nasal/diagnóstico , Pruebas de Provocación Nasal , Sustancia P , Tiorfan/farmacología , Tiorfan/uso terapéuticoRESUMEN
Neutral endopeptidase (NEP) (enkephalinase, EC 3.4.24.11) and angiotensin converting enzyme (ACE) are two peptidases that can cleave the C-terminal dipeptide bradykinin(8-9) from bradykinin. To determine whether these peptidases play roles in modulating kinin-induced contractions in the airways, we studied the effects of captopril, an ACE inhibitor, and of leucine-thiorphan and phosphoramidon, two NEP inhibitors, on the contractile responses to bradykinin and lysyl-bradykinin in isolated segments of ferret trachea. Bradykinin and lysyl-bradykinin-induced contractions in a concentration-dependent fashion (P less than .001), with a threshold of 10(-7) M and 5 x 10(-7) M, respectively. In contrast, the bradykinin(8-9) and the N-terminal heptapeptide bradykinin(1-7), the major fragments of hydrolysis of bradykinin by NEP and ACE, had a very weak or no effect on tracheal contraction in concentrations as great as 10(-5) M. Captopril, leucine-thiorphan and phosphoramidon (each inhibitor at 10(-5) M, 15 min) shifted the concentration-response curves to lower concentrations by approximately 1 to 1.5 log U (P less than .05). Both NEP inhibitors and the ACE inhibitor potentiated the response to bradykinin in a concentration-dependent fashion (P = .0001), and the combination of phosphoramidon and captopril resulted in an additive potentiation of bradykinin-induced contraction (P less than .02). [D-Pro2-D-Trp7,9]-substance P, a substance P antagonist, did not modify the potentiation of bradykinin-induced contraction by NEP inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Cininas/farmacología , Metaloendopeptidasas/antagonistas & inhibidores , Contracción Muscular/efectos de los fármacos , Tiorfan/análogos & derivados , Tráquea/efectos de los fármacos , Animales , Captopril/farmacología , Relación Dosis-Respuesta a Droga , Sinergismo Farmacológico , Hurones , Glicopéptidos/farmacología , Masculino , Neprilisina , Tiopronina/análogos & derivados , Tiopronina/farmacologíaRESUMEN
Acute exposure to cigarette smoke provokes airway hyperresponsiveness to substance P and inactivates neutral endopeptidase (NEP). To determine whether nedocromil sodium can prevent cigarette smoke-induced hyperresponsiveness to substance P, we studied two groups of anaesthetized guinea-pigs. One group of guinea-pigs was pretreated with aerosolized 0.9% NaCl solution (90 breaths), the other group was pretreated with aerosolized nedocromil sodium (10(-4) M, 90 breaths). In each animal, pretreatment was followed by either exposure to the smoke of one cigarette or exposure to air. After acute exposure to cigarette smoke or to air, we measured the change in total pulmonary resistance (RL) induced by increasing concentrations of aerosolized substance P. In the absence of nedocromil sodium, the bronchoconstrictor responses to substance P were greater in cigarette smoke-exposed guinea-pigs than in air-exposed animals. Aerosolized nedocromil sodium had no effect on the response to substance P in air-exposed animals, but it reduced cigarette smoke-induced hyperresponsiveness to substance P. The preventive effect on cigarette smoke-induced hyperresponsiveness to substance P was observed at concentrations of aerosolized nedocromil sodium of 3 x 10(-5), 10(-4), and 3 x 10(-4) M. In vitro, cigarette smoke solution inhibited NEP activity from lung membrane preparations, but this inhibitory effect was not modified by nedocromil sodium (10(-4) M). We conclude that aerosolized nedocromil sodium reduces cigarette smoke-induced airway hyperresponsiveness to substance P in vivo. This action of nedocromil sodium is not due to a protective effect on cigarette smoke-induced inactivation of NEP in vitro.
Asunto(s)
Broncoconstricción/efectos de los fármacos , Nedocromil/farmacología , Sustancia P/farmacología , Contaminación por Humo de Tabaco , Aerosoles , Animales , Hiperreactividad Bronquial/inducido químicamente , Hiperreactividad Bronquial/fisiopatología , Broncoconstricción/fisiología , Relación Dosis-Respuesta a Droga , Cobayas , Técnicas In Vitro , Pulmón/enzimología , Masculino , Neprilisina/metabolismoRESUMEN
Heat shock proteins (HSPs), which are important targets for gammadelta T-lymphocytes, are thought to play a role in inflammatory and immune diseases. The purpose of this study was to characterize, in asthma, the presence and distribution of alphabeta and gammadelta T-lymphocytes and of hsp60, hsp70 and hsp90 in bronchial biopsies, and to seek for a co-localization of gammadelta T-cells and HSPs. Ten subjects with mild atopic asthma and nine control subjects underwent fibreoptic bronchoscopy and bronchial biopsies, to which specific monoclonal antibodies and immunohistochemical techniques were applied. T-lymphocytes present in bronchi both of asthmatic and control subjects were predominantly of the alphabeta T-cell receptor phenotype (median 642 cells x mm(-2) (range 85-1,510 cells x mm(-2)), and 855 cells x mm(-2) (286-2,424 cells x mm(-2)), respectively), whereas, gammadelta T-lymphocytes were always rare (median 26 cells x mm(-2) (range 0-114 cells x mm(-2)), and 0 cells x mm(-2 (0-57 cells x mm(-2), respectively). Both in asthmatic and control subjects, bronchial epithelium was positive for hsp60, hsp70 and hsp90. There was no significant difference in the percentages of positive epithelial cells between asthmatic and control subjects. No co-localization of HSPs and gammadelta T-cells was observed. Our findings do not support the hypothesis that heat shock proteins and gammadelta T-cells play an important role in inflammatory and immune responses in mild asthma.
Asunto(s)
Asma/inmunología , Bronquios/metabolismo , Proteínas de Choque Térmico/biosíntesis , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Linfocitos T/inmunología , Adulto , Asma/metabolismo , Asma/patología , Biopsia , Bronquios/patología , Broncoscopía , Estudios de Casos y Controles , Femenino , Humanos , Técnicas para Inmunoenzimas , Masculino , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Linfocitos T/metabolismoRESUMEN
The goals of the long-term management of reversible obstructive airways disease (ROAD) are to find the minimum treatment that controls symptoms, allows resumption of normal life, prevents severe attack and death, and controls airflow obstruction. ROADs include asthma, chronic bronchitis, and emphysema. Although the differential diagnosis between these different entities may be difficult, they share the same possibilities of pharmacotherapy, including bronchodilator and antiinflammatory drugs. beta 2-agonists administered via inhaled route produce the best bronchodilator/side effects ratio, provided that the drugs reach the bronchi. This underlines the importance of a proper inhalation technique when using a metered-dose inhaler. In patients with hand-breath coordination problems, powder inhalers or spacer devices are useful to ameliorate the therapeutic efficacy of inhaled drugs. Anticholinergic agents are usually less potent bronchodilators than inhaled beta 2 agonists in asthma, but they may have additive effects when associated with beta 2 agonists. Only a therapeutic trial with peak-flow monitoring can demonstrate the efficacy of anticholinergic drugs in individuals. Theophylline's kinetics are characterized by a narrow therapeutic index with high inter- and intraindividual variabilities. Sodium cromoglycate and nedocromil sodium are antiallergic drugs, the efficacy of which has been demonstrated in controlled studies. Corticosteroids are the most efficient anti-asthma drugs. Inhaled corticosteroid dosing should be tailored to each individual. If inhaled corticosteroid therapy is used in an oral corticosparing attempt, patients should be followed-up during several months. The management of ROAD includes the diagnostic procedures, the identification of triggers and inducers of airways obstruction, the assessment of severity of the disease, and then the treatment and education of the patient. Strategy design to achieve proper use of drugs by patients is discussed.
Asunto(s)
Antiinflamatorios/uso terapéutico , Broncodilatadores/uso terapéutico , Enfermedades Pulmonares Obstructivas/tratamiento farmacológico , Agonistas Adrenérgicos beta/uso terapéutico , Resistencia de las Vías Respiratorias/efectos de los fármacos , Niño , Humanos , Cuidados a Largo PlazoRESUMEN
BACKGROUND: An easy and reliable method to measure potential difference (PD) in the lower airways would be of interest in the field of cystic fibrosis. We have developed silver/silver chloride (Ag/AgCl) electrodes to measure PD in the lower airways. METHODS: To validate this technique the nasal PD measured with Ag/AgCl electrodes and with conventional perfused electrodes was compared in 16 patients. The range of PD measured with Ag/AgCl electrodes in the lower airways during fibreoptic bronchoscopy was determined in 14 adult patients and in nine the reproducibility of this technique was examined. RESULTS: Nasal PD values measured with Ag/AgCl and perfused electrodes were highly correlated (r = 0.985, p < 0.0001) and the limits of agreement (mean +/- 2SD of the difference) between the two methods were -1.91 mV and 1.53 mV. In the lower airways a progressive and slight decrease in PD values with decreasing airway diameter was observed in most patients. The mean (2SD) of the differences between the two tracheal measurements was 0.21 (1.73) mV. CONCLUSIONS: The use of Ag/AgCl electrodes gives a reliable and reproducible measurement of PD in the lower airways in humans.