RESUMO
BACKGROUND: Exposure to environmental pollutants promotes Th2 cell responses. Aryl hydrocarbon receptor (AhR) activation aggravates allergic responses. Epithelium-derived thymic stromal lymphopoietin (TSLP), interleukin (IL)-25, and IL-33 are implicated in the dysregulation of Th2 immune responses in severe allergic asthma. METHODS: Bronchial biopsies of 28 allergic severe asthma and 6 mild asthma subjects from highly polluted areas were analyzed for AhR nuclear translocation (NT), cytokine expression, and gene activation. Cultured primary epithelial cells were stimulated with diesel exhausted particles (DEP) to determine AhR-mediated IL-33, Il-25, and TSLP synthesis and release. RESULTS: Primary bronchial epithelial cells exposed to DEP showed upregulation of IL-33, IL-25, and TSLP. These effects were abolished by knockdown of AhR by siRNA. Increased AhR/ARNT binding to promoters of IL-33, IL-25, and TSLP was found using chromatin immunoprecipitation (ChIP) assay. Allergic severe asthma with high AhR NT had higher bronchial gene and protein expression of IL-33, IL-25, and TSLP. These patients derived clinical benefit from anti-IgE treatment. CONCLUSION: Aryl hydrocarbon receptor activation by DEP mediates upregulation of IL-33, IL-25, and TSLP with Th2 activation, potentially linking environmental pollution and allergic severe asthma.
Assuntos
Asma/etiologia , Asma/metabolismo , Citocinas/metabolismo , Receptores de Hidrocarboneto Arílico/metabolismo , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Emissões de Veículos , Alérgenos/imunologia , Anticorpos Anti-Idiotípicos/farmacologia , Anticorpos Anti-Idiotípicos/uso terapêutico , Asma/diagnóstico , Asma/terapia , Biópsia , Citocinas/genética , Feminino , Imunofluorescência , Regulação da Expressão Gênica , Humanos , Imunoglobulina E/imunologia , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Regiões Promotoras Genéticas , Transporte Proteico , Testes de Função Respiratória , Mucosa Respiratória/patologia , Células Th2/imunologia , Células Th2/metabolismo , Linfopoietina do Estroma do TimoRESUMO
BACKGROUND: Bone metastases are common in patients with advanced non-small-cell lung cancer (NSCLC) and can have devastating consequences. Preventing or delaying bone metastases may improve outcomes. PATIENTS AND METHODS: This study evaluated whether zoledronic acid (ZOL) delayed disease progression or recurrence in patients with controlled stage IIIA/B NSCLC after first-line therapy. Patients received vitamin D and calcium supplementation and were randomized to i.v. ZOL (every 3-4 weeks) or no treatment (control). The primary end point was progression-free survival (PFS). RESULTS: No significant intergroup differences were observed in PFS or overall survival (OS). Median PFS was 9.0 months with ZOL versus 11.3 months for control. Fifteen ZOL-treated (6.6%) and 19 control patients (9.0%) developed bone metastases. Estimated 1-year OS was 81.8% for each group. ZOL safety profile was consistent with previous clinical data, but with higher discontinuations versus control. Fifteen ZOL-treated (6.6%) and five control patients (2.3%) had renal adverse events. Two cases of osteonecrosis of the jaw were reported. CONCLUSIONS: ZOL did not significantly affect PFS or OS in stage IIIA/B NSCLC patients with controlled disease, with a trend toward worsening PFS in the longer-term follow-up. Few patients experienced bone metastases, possibly limiting the potential ZOL impact on disease course.
Assuntos
Neoplasias Ósseas/prevenção & controle , Neoplasias Ósseas/secundário , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Difosfonatos/uso terapêutico , Imidazóis/uso terapêutico , Neoplasias Pulmonares/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Cálcio/administração & dosagem , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma Pulmonar de Células não Pequenas/secundário , Suplementos Nutricionais , Feminino , Humanos , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Vitamina D/administração & dosagem , Adulto Jovem , Ácido ZoledrônicoRESUMO
The self-management of asthma can improve clinical outcomes. Recently, mobile telephones have been widely used as an efficient, instant personal communication tool. This study investigated whether a self-care system will achieve better asthma control through a mobile telephone-based interactive programme. This was a prospective, controlled study in outpatient clinics. From 120 consecutive patients with moderate-to-severe persistent asthma, 89 were eventually recruited for the study, with 43 in the mobile telephone group (with a mobile telephone-based interactive asthma self-care system). In the mobile telephone group, mean ± sem peak expiratory flow rate significantly increased at 4 (378.2 ± 9.3 L·min⻹; n = 43; p = 0.020), 5 (378.2 ± 9.2 L·min⻹; n = 43; p = 0.008) and 6 months (382.7 ± 8.6 L·min⻹; n = 43; p = 0.001) compared to the control group. Mean±sem forced expiratory volume in 1 s significantly increased at 6 months (65.2 ± 3.2% predicted; n = 43; p < 0.05). Patients in the mobile telephone group had better quality of life after 3 months, as determined using the Short Form-12® physical component score, and fewer episodes of exacerbation and unscheduled visits than the control group. Patients in the mobile telephone group significantly increased their mean daily dose of either systemic or inhaled corticosteroids compared with the control group. The mobile telephone-based interactive self-care system provides a convenient and practical self-monitoring and -management of asthma, and improves asthma control.
Assuntos
Corticosteroides/uso terapêutico , Antiasmáticos/uso terapêutico , Asma/tratamento farmacológico , Telefone Celular , Autocuidado/métodos , Feminino , Humanos , Antagonistas de Leucotrienos/uso terapêutico , Masculino , Pessoa de Meia-Idade , Qualidade de Vida , Testes de Função Respiratória , Telemedicina , Resultado do TratamentoRESUMO
BACKGROUND: Identification of appropriate markers for predicting clinical benefit with erlotinib in non-small-cell lung cancer (NSCLC) may be able to guide patient selection for treatment. This open-label, multicentre, phase II trial aimed to identify genes with potential use as biomarkers for clinical benefit from erlotinib therapy. METHODS: Adults with stage IIIb/IV NSCLC in whom one or more chemotherapy regimen had failed were treated with erlotinib (150 mg/day). Tumour biopsies were analysed using gene expression profiling with Affymetrix GeneChip microarrays. Differentially expressed genes were verified using quantitative RT-PCR (qRT-PCR). RESULTS: A total of 264 patients were enrolled in the study. Gene expression profiles found no statistically significant differentially expressed genes between patients with and without clinical benefit. In an exploratory analysis in responding versus nonresponding patients, three genes on chromosome 7 were expressed at higher levels in the responding group [epidermal growth factor receptor (EGFR), phosphoserine phosphatase (PSPH) and Rap guanine nucleotide exchange factor 5 (RAPGEF5)]. Independent quantification using qRT-PCR validated the association between EGFR and PSPH overexpression, but not RAPGEF5 overexpression, and clinical outcome. CONCLUSIONS: This study supports the use of erlotinib as an alternative to chemotherapy for patients with relapsed advanced NSCLC. Genetic amplification of the EGFR region of chromosome 7 may be associated with response to erlotinib therapy.
Assuntos
Biomarcadores Tumorais/genética , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Perfilação da Expressão Gênica , Neoplasias Pulmonares/tratamento farmacológico , Quinazolinas/uso terapêutico , Adulto , Idoso , Idoso de 80 Anos ou mais , Antineoplásicos/efeitos adversos , Antineoplásicos/uso terapêutico , Biomarcadores Farmacológicos/análise , Biomarcadores Farmacológicos/metabolismo , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/genética , Intervalo Livre de Doença , Cloridrato de Erlotinib , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , Masculino , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , Prognóstico , Quinazolinas/efeitos adversosRESUMO
BACKGROUND: This retrospective study aimed to determine the factors leading to obstructive granulation tissue formation after the placement of a self-expandable metallic stent (SEMS) in patients with benign tracheal disease. METHODS: From 2001 to 2007, a total of 67 patients (age: 62.1 +/- 15.4 years; range: 23-87 years) with benign tracheal disease received 75 ultraflex SEMS in our institution. RESULTS: There were 35 SEMSs complicated by obstructive granulation tissue formation out of the 75 stents placed in patients with tracheal disease, giving an incidence of 47.8 % (32/67 patients). The median time until developing granulation tissue was 106 days (IQR, 46-396). Structural airway obstruction prior to SEMS implantation independently predicted obstructive granulation tissue formation after SEMS implantation (odds ratio: 3.84; 95 % CI: 1.01-8.7; P = 0.04). Time to granulation tissue detection was shorter in patients with structural airway obstruction before SEMS implantation (structural airway obstruction vs. dynamic collapse airway: median [IQR] 95 [38-224, n = 26] vs. 396 days [73-994, n = 9]; P = 0.02). CONCLUSIONS: Obstructive granulation tissue formation is not uncommon after SEMS implantation and structural airway obstruction prior to SEMS implantation is an independent predictor. Although SEMS implantation should be restricted to a select population, it may be placed in patients not suitable for surgical intervention or rigid bronchoscopy with anesthesia because of poor pulmonary function.
Assuntos
Obstrução das Vias Respiratórias/etiologia , Reação a Corpo Estranho/etiologia , Tecido de Granulação/patologia , Stents/efeitos adversos , Estenose Traqueal/terapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Obstrução das Vias Respiratórias/diagnóstico , Obstrução das Vias Respiratórias/patologia , Obstrução das Vias Respiratórias/terapia , Broncoscopia , Reação a Corpo Estranho/diagnóstico , Reação a Corpo Estranho/patologia , Reação a Corpo Estranho/terapia , Humanos , Estimativa de Kaplan-Meier , Modelos Logísticos , Pessoa de Meia-Idade , Razão de Chances , Seleção de Pacientes , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Fatores de Tempo , Tomografia Computadorizada por Raios X , Estenose Traqueal/diagnóstico , Adulto JovemRESUMO
OBJECTIVES: To determine predictive factors for mortality among pulmonary tuberculosis (PTB) patients without human immunodeficiency virus (HIV) infection and in need of mechanical ventilation (TBMV). METHODS: From July 2004 to December 2005, 612 respiratory failure patients requiring mechanical ventilation were admitted to the intensive care unit (ICU) of Chang Gung Memorial Hospital, Taipei, Taiwan. Of these, 59 non-HIV-infected patients had active PTB as the primary cause. Mortality rates were measured in TBMV patients and predictors were investigated. Incidence of treatment delay for nosocomial pneumonia was compared between survivors and fatalities. RESULTS: Of the 59 patients with TBMV, 40 (67.8%) died in the ICU. Multi-organ failure syndrome (OR 8.59, 95%CI 1.85-101.27) and nosocomial pneumonia (OR 5.77, 95%CI 1.33-44.36) were independently associated with in-hospital mortality. Treatment delay >24 h for nosocomial pneumonia was significantly more frequent among fatalities than among survivors (19/26, 73.1% vs. 0/3, 0%; P = 0.033). CONCLUSION: Nosocomial pneumonia in TB patients with respiratory failure is associated with a poor prognosis; this appears to be further aggravated by delays in appropriate treatment. Measures to prevent nosocomial pneumonia should be carefully instituted and treatment for nosocomial pneumonia should be started promptly among such patients.
Assuntos
Insuficiência Respiratória/mortalidade , Tuberculose Pulmonar/mortalidade , APACHE , Idoso , Idoso de 80 Anos ou mais , Infecção Hospitalar/mortalidade , Humanos , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Respiração Artificial , Insuficiência Respiratória/etiologia , Insuficiência Respiratória/terapia , Estudos Retrospectivos , Tuberculose Pulmonar/complicaçõesRESUMO
Stent implantation has been reported to facilitate liberation from mechanical ventilation in patients with respiratory failure due to central airway disease. The present retrospective cohort study sought to evaluate the risk and benefit of stent implantation via bronchoscopy without fluoroscopic guidance in mechanically ventilated patients. From July 2001 to September 2006, 26 patients with acute respiratory failure were recruited. A bronchoscope was inserted through a mouth guard into the space between the tracheal wall and the endotracheal tube. A guide wire was inserted via the flexible bronchoscope to the lesion site. The bronchoscope was reintroduced through the endotracheal tube. Under bronchoscopic visualisation, the delivery catheter was advanced over the guide wire to deploy the stent. These procedures were successfully performed in 26 patients, with 22 stents placed in the trachea and seven in the main bronchus. Of the 26 patients, 14 (53.8%) became ventilator independent during their stay in the intensive care unit. Severe pneumonia was the most common cause, in seven (58.3%) out of 12 patients, for continued ventilator dependence after stenting. Granulation tissue formation was found in seven patients during the follow-up period. It is concluded that metallic stents can be safely implanted without fluoroscopic guidance in patients with respiratory failure, to facilitate ventilator independence.
Assuntos
Obstrução das Vias Respiratórias/cirurgia , Broncoscopia/métodos , Insuficiência Respiratória/cirurgia , Stents , Idoso , Obstrução das Vias Respiratórias/complicações , Broncoscopia/efeitos adversos , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Respiração Artificial , Insuficiência Respiratória/etiologia , Estudos Retrospectivos , Desmame do RespiradorRESUMO
The application of a supervised endurance exercise training programme in a home setting offering convenience and prolonged effects is a challenge. In total, 48 patients were initially assessed by the incremental shuttle walk test (ISWT), spirometry and the Short Form-12 (SF-12) quality-of-life questionnaire, and then every 4 weeks for 3 months thereafter and again after 1 yr. During the first 3 months, 24 patients in the cell phone group were asked to perform daily endurance walking at 80% of their maximal capacity by following the tempo of music from a program installed on a cell phone. The level of endurance walking at home was readjusted monthly according to the result of ISWT. In the control group, 24 patients received the same protocol and were verbally asked to take daily walking exercise at home. Patients in the cell phone group significantly improved their ISWT distance and duration of endurance walking after 8 weeks. The improvements in ISWT distance, inspiratory capacity and SF-12 scoring at 12 weeks persisted until the end of the study, with less acute exacerbations and hospitalisations. In the present pilot study, the cell phone-based system provides an efficient, home endurance exercise training programme with good compliance and clinical outcomes in patients with moderate-to-severe chronic obstructive pulmonary disease.
Assuntos
Telefone Celular , Terapia por Exercício/métodos , Doença Pulmonar Obstrutiva Crônica/reabilitação , Idoso , Tolerância ao Exercício , Feminino , Humanos , Masculino , Música , Cooperação do Paciente , Projetos Piloto , Qualidade de VidaRESUMO
BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a heterogeneous condition that can differ in its clinical manifestation, structural changes and response to treatment. OBJECTIVE: To identify subgroups of COPD with distinct phenotypes, evaluate the distribution of phenotypes in four related regions and calculate the 1-year change in lung function and quality of life according to subgroup. METHODS: Using clinical characteristics, we performed factor analysis and hierarchical cluster analysis in a cohort of 1676 COPD patients from 13 Asian cities. We compared the 1-year change in forced expiratory volume in one second (FEV1), modified Medical Research Council dyspnoea scale score, St George's Respiratory Questionnaire (SGRQ) score and exacerbations according to subgroup derived from cluster analysis. RESULTS: Factor analysis revealed that body mass index, Charlson comorbidity index, SGRQ total score and FEV1 were principal factors. Using these four factors, cluster analysis identified three distinct subgroups with differing disease severity and symptoms. Among the three subgroups, patients in subgroup 2 (severe disease and more symptoms) had the most frequent exacerbations, most rapid FEV1 decline and greatest decline in SGRQ total score. CONCLUSION: Three subgroups with differing severities and symptoms were identified in Asian COPD subjects.
Assuntos
Dispneia/epidemiologia , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Qualidade de Vida , Idoso , Ásia/epidemiologia , Cidades , Análise por Conglomerados , Estudos de Coortes , Dispneia/etiologia , Análise Fatorial , Feminino , Seguimentos , Volume Expiratório Forçado , Humanos , Masculino , Pessoa de Meia-Idade , Fenótipo , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Fatores de Risco , Índice de Gravidade de Doença , Inquéritos e QuestionáriosRESUMO
To ensure the success of systemic gene therapy, it is critical to enhance the tumor specificity and activity of the promoter. In the current study, we determined that topoisomerase IIalpha promoter is selectively activated in breast cancer cells. An element containing an inverted CCAAT box (ICB) was shown to be responsible for the breast cancer specificity. When the ICB-harboring topoisomerase IIalpha minimal promoter was linked with an enhancer sequence from the cytomegalovirus immediate early gene promoter (CMV promoter), this composite promoter, CT90, exhibited activity comparable to or higher than the CMV promoter in breast cancer cells in vitro and in vivo, yet expresses much lower activity in normal cell lines and normal organs than the CMV promoter. A CT90-driven construct expressing BikDD, a potent proapoptotic gene, was shown to selectively kill breast cancer cells in vitro, and to suppress mammary tumor development in an animal model of intravenously administrated, liposome-delivered gene therapy. Expression of BikDD was readily detectable in the tumors but not in the normal organs (such as heart) of CT90-BikDD-treated animals. The results indicate that liposomal CT90-BikDD is an effective systemic breast cancer-targeting gene therapy.
Assuntos
Antígenos de Neoplasias/genética , Proteínas Reguladoras de Apoptose/genética , Neoplasias da Mama/terapia , DNA Topoisomerases Tipo II/genética , Proteínas de Ligação a DNA/genética , Terapia Genética , Proteínas de Membrana/genética , Mutação , Elementos de Resposta/genética , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Citomegalovirus/genética , Elementos Facilitadores Genéticos/genética , Feminino , Expressão Gênica , Vetores Genéticos , Humanos , Lipossomos , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Nus , Proteínas Mitocondriais , Neoplasias ExperimentaisRESUMO
1. Goblet cell secretion in guinea-pig airways is under neural control. Opioids have previously been shown to inhibit neurogenic plasma exudation and bronchoconstriction in guinea-pig airways. We have now examined the effects of morphine and opioid peptides on tracheal goblet cell secretion induced by either electrical stimulation of the cervical vagus nerves, exogenous capsaicin, or acute inhalation of cigarette smoke. The degree of goblet cell secretion was determined by a morphometric method and expressed as a mucus score which is inversely related to mucus discharge. 2. Morphine, 1 mg kg-1, completely blocked goblet cell secretion induced by electrical stimulation of the vagus nerves. Morphine also inhibited the response to cigarette smoke given either at a low dose (10 breaths of 1:10 diluted in air), which principally activates cholinergic nerves, or at a high dose (20 breaths of undiluted), which activates capsaicin-sensitive sensory nerves, by 100% and 73% respectively. In contrast, morphine had no significant inhibitory effect on capsaicin-induced goblet cell secretion. The inhibitory effect of morphine was reversed by naloxone. 3. Selective mu- or delta-opioid receptor agonists, [D-Ala2, NMePhe4, Glyol5]enkephalin (DAMGO) or [D-Pen2, D-Pen5]enkephalin (DPDPE) respectively, caused a dose-related inhibition of low dose cigarette smoke-induced goblet cell discharge, with DPDPE more potent than DAMGO. A kappa-receptor agonist, trans-3,4-dichloro-N-methyl-N-(2-(1-pyrollidinyl)cyclohexyl) benzeneacetamine (U-50,488H), had no inhibitory effect. DPDPE had no inhibitory effect on goblet cell secretion induced by exogenous methacholine. 4. DAMGO dose-dependently blocked the response to high dose cigarette smoke with a maximal inhibition of 95% at 2 x 10(-7) mol kg-1. Neither DPDPE nor U-50,488H had any significant inhibitory effect. The increase in goblet cell secretion induced by exogenous substance P was not affected by DAMGO.5. We conclude that opioids inhibit neurally-mediated goblet cell secretion via actions at prejunctional delta and mu-receptors on cholinergic nerves and at mu-receptors on sensory nerve endings, and that capsaicin activation of sensory nerves is via a different mechanism from that of electrical or cigarette smoke activation.
Assuntos
Capsaicina/farmacologia , Entorpecentes/farmacologia , Fumar/metabolismo , Traqueia/metabolismo , Animais , Estimulação Elétrica , Cobaias , Técnicas In Vitro , Masculino , Compostos de Metacolina/farmacologia , Morfina/farmacologia , Receptores Opioides/efeitos dos fármacos , Receptores Opioides delta , Receptores Opioides mu , Substância P/farmacologia , Traqueia/citologia , Traqueia/efeitos dos fármacos , Nervo Vago/efeitos dos fármacos , Nervo Vago/fisiologiaRESUMO
1. Tumour necrosis factor-alpha (TNF-alpha) is implicated in the pathogenesis of many pulmonary and airway diseases. TNF-alpha stimulation may release interleukin-8 (IL-8) in airways mediated via an increase in intracellular oxidant stress. In the present study, we have assessed leukosequestration and IL-8 release in the airways in response to intratracheal administration of human recombinant TNF-alpha, and examined the modulatory role of endogenous NO by pretreatment with a NO synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME). 2. TNF-alpha (10(2)-10(-4) u) was administered intratracheally in male guinea-pigs which were anaesthetized with urethane and were ventilated artificially. TNF-alpha induced a time- and dose-related increase in neutrophil numbers and a concomitant increase in human IL-8 equivalent level retrieved from bronchoalveolar lavage (BAL) with the peak effect at 10(3) u at 6 h of TNF-alpha injection (late phase). Intratracheal administration of recombinant human (rh)IL-8 (0.025, 0.25, 2.5 ng) producing a similar range of human IL-8 equivalent levels in BAL as measured in our results induced neutrophil recovery in BAL fluid to a similar extent. Administration of anti-IL-8 antibody prevented the late phase of neutrophil recruitment induced by TNF-alpha or rhIL-8. 3. Pretreatment with L-NAME significantly enhanced the TNF-alpha (10(3) u)-induced neutrophil recruitment and human IL-8 equivalents production at 6 h, but not at 1 h of TNF-alpha administration (early phase). L-Arginine reversed the responses to L-NAME. Pretreatment with 0.2% DMSO (i.v.) significantly inhibited TNF-alpha-induced neutrophil recruitment and human IL-8 equivalents release both in the early and late phase of the responses. Pretreatment with DMSO also inhibited the enhancement effect of L-NAME on the late phase of TNF-alpha-induced responses. DMSO failed to modify exogenous rhIL-8-induced neutrophil recruitment. Neither L-NAME nor DMSO alone induced any significant change in neutrophil numbers or human IL-8 equivalent level in BAL fluid. 4. Neutrophil depletion by cyclophosphamide pretreatment failed to modify TNF-alpha-induced human IL-8 equivalent release. 5. The expression of beta 2-integrin, CD11b/CD18 on neutrophils was increased only in the late but not early phase of TNF-alpha stimulation. L-NAME failed to modify these responses. 6. In conclusion, we demonstrated that NO may be an important endogenous inhibitor of TNF-alpha-induced leukocyte chemotaxis via inhibition of IL-8 production. Thus, the production of NO in airway inflammatory diseases may play a negative feedback role in self-limiting the magnitude of inflammatory responses.
Assuntos
Interleucina-8/metabolismo , Neutrófilos/efeitos dos fármacos , Óxido Nítrico/fisiologia , Traqueia/efeitos dos fármacos , Traqueia/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Animais , Pressão Sanguínea/efeitos dos fármacos , Líquido da Lavagem Broncoalveolar/química , Antígenos CD18/biossíntese , Ciclofosfamida/farmacologia , Interações Medicamentosas , Inibidores Enzimáticos/farmacologia , Cobaias , Humanos , Imunossupressores/farmacologia , Antígeno de Macrófago 1/biossíntese , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Neutrófilos/citologia , Neutrófilos/metabolismo , Óxido Nítrico/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Proteínas Recombinantes/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
1. There are increased numbers of circulating CD34(+) progenitor cells for eosinophils in patients with atopic asthma, with a further increase following allergen exposure or spontaneous worsening of asthma. We investigated the expression of IL-5 and IL-5Ralpha receptor in circulating CD34(+) progenitor cells in allergic asthmatics and the effects of corticosteroids. 2. Using double-staining techniques, up to 50% of CD34(+) cells expressed intracellular IL-5, and by RT - PCR, there was significant expression of IL-5 mRNA. When cultured in a semi-liquid methylcellulose medium, there were more eosinophil colony-forming units grown from asthmatic non-adherent mononuclear cell depleted of T cells in the presence of the growth factors GM-CSF, SCF and IL-3, but not of IL-5. 3. An anti-IL-5Ralpha receptor antibody and an anti-sense IL-5 oligonucleotide reduced the number of eosinophil colony forming units. No IL-5 mRNA or protein expression on T cells was observed in asthmatics or normal subjects. In the presence of growth factors including IL-5, there were significantly greater colony numbers with eosinophilic lineage grown from either asthmatics or normal subjects. 4. Dexamethasone (10(-6) M) suppressed IL-5 mRNA and protein expression in CD34(+) cells, and reduced eosinophil colony-forming units in asthmatics, but not in normal subjects. Dexamethasone did not change the expression of IL-5Ralpha on CD34(+) cells. 5. We conclude that there is increased expression of IL-5 on blood CD34(+) cells of patients with asthma and that this expression may auto-regulate eosinophilic colony formation from these progenitor cells. Corticosteroids inhibit the expression of IL-5 in circulating CD34(+) progenitor cells.
Assuntos
Asma/fisiopatologia , Diferenciação Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Eosinófilos/efeitos dos fármacos , Interleucina-5/farmacologia , Adulto , Anticorpos Monoclonais/farmacologia , Antígenos CD34/imunologia , Asma/patologia , DNA Antissenso/farmacologia , Dexametasona/farmacologia , Eosinófilos/citologia , Eosinófilos/imunologia , Feminino , Expressão Gênica , Glucocorticoides/farmacologia , Humanos , Interleucina-5/genética , Interleucina-5/fisiologia , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Interleucina/imunologia , Receptores de Interleucina-5 , Células-Tronco/efeitos dos fármacosRESUMO
1. This study investigated the role of protein kinase C (PKC) and transcription factor nuclear factor-kappaB (NF-kappaB) in cyclooxygenase-2 (COX-2) expression caused by lipoteichoic acid (LTA), a cell wall component of the gram-positive bacterium Staphylococcus aureus, in human pulmonary epithelial cell line (A549). 2. LTA caused dose- and time-dependent increases in COX-2 expression and COX activity, and a dose-dependent increase in PGE(2) release in A549 cells. The LTA-induced increases in COX-2 expression and COX activity were markedly inhibited by dexamethasone, actinomycin D or cyclohexamide, but not by polymyxin B, which binds and inactivates endotoxin. 3. The phosphatidylcholine-phospholipase C (PC-PLC) inhibitor (D-609) and the phosphatidate phosphohydrolase inhibitor (propranolol) reduced the LTA-induced increases in COX-2 expression and COX activity, while phosphatidylinositol-phospholipase C inhibitor (U-73122) had no effect. The PKC inhibitors (Go 6976, Ro 31-8220 and GF 109203X) and NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), also attenuated the LTA-induced increases in COX-2 expression and COX activity. 4. Treatment of A549 cells with LTA caused an increase in PKC activity in the plasma membrane; this stimulatory effect was inhibited by D-609, propranolol, or Go 6976, but not by U-73122. 5. Exposure of A549 cells to LTA caused a translocation of p65 NF-kappaB from the cytosol to the nucleus and a degradation of IkappaB-alpha in the cytosol. Treatment of A549 cells with LTA caused NF-kappaB activation by detecting the formation of NF-kappaB-specific DNA-protein complex in the nucleus; this effect was inhibited by dexamethasone, D-609, propranolol, Go 6976, Ro 31-8220, or PDTC. 6. These results suggest that LTA might activate PC-PLC and phosphatidylcholine-phospholipase D to induce PKC activation, which in turn initiates NF-kappaB activation, and finally induces COX-2 expression and PGE(2) release in human pulmonary epithelial cell line.
Assuntos
Células Epiteliais/efeitos dos fármacos , Células Epiteliais/enzimologia , Isoenzimas/biossíntese , Lipopolissacarídeos/farmacologia , Pulmão/enzimologia , NF-kappa B/biossíntese , Prostaglandina-Endoperóxido Sintases/biossíntese , Transdução de Sinais/efeitos dos fármacos , Staphylococcus aureus , Ácidos Teicoicos/farmacologia , Ciclo-Oxigenase 2 , Dinoprostona/biossíntese , Relação Dose-Resposta a Droga , Células Epiteliais/microbiologia , Humanos , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/fisiologia , Proteínas de Membrana , Transdução de Sinais/fisiologia , Células Tumorais CultivadasRESUMO
1. Lipopolysaccharide (LPS) is implicated in many pulmonary and airway inflammatory diseases. Tachykinins released from nerve endings increase vascular permeability. In this study, we have assessed the enhancement by LPS of tachykinin-mediated plasma exudation in guinea-pig airways, and examined the role of oxidants as well as leukocyte adherence. 2. LPS (100 microg kg(-1), i.v.) was administered 0-3 h before bilateral electrical stimulation of the cervical vagus nerves in animals anaesthetized with urethane and ventilated. Vagal stimulation increased vascular permeability in the airways. LPS enhanced the vagally-mediated plasma exudation with the peak effect at 1 h after LPS administration. LPS alone induced no significant plasma exudation. LPS also enhanced exogenous substance P (10(-8) mol kg(-1), i.v.)-induced plasma exudation. 3. The NK-1 receptor antagonist L-732,138 abolished vagally-induced plasma exudation and significantly inhibited the enhancement by LPS. Pretreatment with superoxide dismutase (SOD, 5000 U kg(-1), i.p.) did not affect the vagally-induced plasma exudation, but inhibited the LPS-enhanced neurogenic plasma leakage. The LPS-enhanced vagally-induced plasma exudation was not completely inhibited by either L-732,138 or SOD pretreatment alone, but was blocked by the combination of both pretreatments. 4. Neutrophil depletion by cyclophosphamide alone did not influence vagally-induced plasma exudation, but significantly inhibited the LPS-enhanced response. 5. In conclusion, we have demonstrated LPS enhanced neurogenic plasma exudation by augmenting the response to tachykinins, partly through NK-1 receptors, to directly increase vascular permeability or to enhance leukocyte adhesion-mediated endothelial cell injury. Tachykinins released from nerve endings may contribute to endotoxin-related airway inflammatory responses.
Assuntos
Permeabilidade Capilar/efeitos dos fármacos , Exsudatos e Transudatos , Lipopolissacarídeos/farmacologia , Terminações Nervosas/metabolismo , Taquicininas/farmacologia , Animais , Relação Dose-Resposta a Droga , Exsudatos e Transudatos/efeitos dos fármacos , Exsudatos e Transudatos/metabolismo , Cobaias , Masculino , Antagonistas dos Receptores de Neurocinina-1 , Neutrófilos/metabolismo , Sistema Nervoso Parassimpático/fisiologia , Organismos Livres de Patógenos Específicos , Superóxido Dismutase/farmacologia , Taquicininas/metabolismo , Triptofano/análogos & derivados , Triptofano/farmacologiaRESUMO
Alveolar macrophages are a heterogeneous cell population. The heterogeneity of alveolar macrophages recovered by bronchoalveolar lavage (BAL) from 12 patients with active pulmonary tuberculosis (TB) and 10 normal subjects was studied using Percoll density fractionation. The numbers and subsets (on the basis of CD3, CD4, and CD8 monoclonal antibodies) of lymphocytes in BAL were measured by flow cytometry. Alveolar macrophages recovered from patients with TB were mainly in the lower-density fractions (< 1.030 and 1.030 to 1.040 g/ml), whereas alveolar macrophages from normal subjects were in the higher-density fractions (1.050 to 1.070 and > 1.070 g/ml). There were no significant differences in alveolar macrophages' repartition between smokers and nonsmokers in either patients with TB or normal subjects. The significant changes in the proportions of the lowest fraction and the higher fractions of alveolar macrophages in patients with TB were not altered after division of our patients into smoker and nonsmoker subgroups when compared with corresponding subgroups in normal subjects. The proportion of the alveolar macrophages in the lowest fraction was inversely related to the bacterial load of sputum and the disease extent on chest radiography in TB patients. The CD4/CD8 ratio was significantly higher in patients with TB. This study shows that alveolar macrophages from TB patients are heterogeneous with hypodense cells predominant probably by interaction with T lymphocytes. Changes in the proportions of alveolar macrophages within subpopulations may be of critical importance in determining the overall response of the lung to TB infection.
Assuntos
Macrófagos Alveolares/classificação , Tuberculose Pulmonar/patologia , Líquido da Lavagem Broncoalveolar/citologia , Relação CD4-CD8 , Contagem de Células , Centrifugação com Gradiente de Concentração , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fumar/patologia , Subpopulações de Linfócitos T , Tuberculose Pulmonar/imunologiaRESUMO
We studied the effect of endogenous nitric oxide (NO) on vagally induced plasma exudation into guinea-pig trachea and main bronchi using 125I-albumin as a plasma marker. NG-Nitro-L-arginine methyl ester (L-NAME, 1-10 mg/kg) dose dependently inhibited neurogenic plasma exudation. Intravenous phenylephrine which simulated the vasopressor effect as L-NAME (10 mg/kg) was without effect. The effect of L-NAME (5 mg/kg) was reversed by L-arginine (50 mg/kg). These results suggest that endogenous NO may contribute to neurogenic inflammation in the airways.
Assuntos
Brônquios/metabolismo , Óxido Nítrico/metabolismo , Plasma/metabolismo , Traqueia/metabolismo , Nervo Vago/fisiologia , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Estimulação Elétrica , Cobaias , Masculino , NG-Nitroarginina Metil ÉsterRESUMO
A potassium (K+) channel activator, BRL 38227, inhibited goblet cell secretion in guinea-pig trachea induced by either electrical stimulation of the vagus nerves or acute inhalation of cigarette smoke, two stimuli which activate both cholinergic nerves and capsaicin-sensitive sensory nerves. BRL 38227 failed to inhibit methacholine- or substance P-induced goblet cell secretion which suggests that K+ channel activators inhibit neurogenic goblet cell secretion via a prejunctional effect on cholinergic and sensory nerves.
Assuntos
Benzopiranos/farmacologia , Broncodilatadores/farmacologia , Canais de Potássio/efeitos dos fármacos , Pirróis/farmacologia , Traqueia/metabolismo , Animais , Cromakalim , Estimulação Elétrica , Cobaias , Masculino , Compostos de Metacolina/antagonistas & inibidores , Compostos de Metacolina/farmacologia , Fumar/fisiopatologia , Substância P/antagonistas & inibidores , Substância P/farmacologia , Traqueia/citologia , Traqueia/efeitos dos fármacos , Nervo Vago/fisiologiaRESUMO
Trimellitic anhydride is a cause of occupational asthma in humans. We have previously found that tracheal instillation of trimellitic anhydride conjugated to guinea pig serum albumin induces acute bronchoconstriction and airway plasma exudation in sensitised animals, responses mediated primarily via histamine release. In the present study, neural mechanisms mediating bronchoconstriction and goblet cell secretion were determined in trimellitic anhydride-sensitised guinea pigs using the ganglionic blocker hexamethonium to eliminate efferent reflex mechanisms, pretreatment with capsaicin to eliminate afferent mechanisms, or cimetidine and mepyramine to eliminate histamine-mediated mechanisms. The magnitude of secretion of intracellular mucus from tracheal goblet cells was quantified morphometrically as a mucus score which is inversely related to the degree of discharge. Guinea pigs were injected intradermally either with 0.1 ml 0.3% trimellitic anhydride in corn oil or with corn oil alone as control. Fourteen to eighteen days later all sensitised animals had developed specific immunoglobulin (Ig) G1 antibodies whereas the controls had not. Tracheal instillation of conjugated trimellitic anhydride in anaesthetised animals significantly increased airway lung resistance (RL) 24-fold in sensitised guinea pigs (34.3 +/- 7.9 cm H2O.ml-1.s) compared with controls (1.4 +/- 0.1 cm H2O.ml-1.s). Mucus score was significantly reduced by 51% (indicating goblet cell secretion) in sensitised guinea pigs (183 +/- 22 mucus score units) compared with controls (372 +/- 41 mucus score units). The antihistamines significantly inhibited conjugated trimellitic anhydride-induced bronchoconstriction by 89%, but did not significantly affect goblet cell discharge. Hexamethonium alone did not significantly affect conjugated trimellitic anhydride-induced bronchoconstriction or goblet cell secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Alérgenos/toxicidade , Broncoconstrição/efeitos dos fármacos , Muco/metabolismo , Anidridos Ftálicos/toxicidade , Traqueia/fisiologia , Resistência das Vias Respiratórias/efeitos dos fármacos , Animais , Permeabilidade Capilar/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Cobaias , Hexametônio/farmacologia , Técnicas In Vitro , Masculino , Exposição Ocupacional , Albumina Sérica/química , Traqueia/citologia , Traqueia/efeitos dos fármacosRESUMO
SETTING: Peripheral blood monocytes (PBM) are the main source of alveolar macrophages, which have an upregulation of inducible nitric oxide synthase (iNOS) in pulmonary tuberculosis (TB). TNF-alpha and IL-1 beta are thought to be involved in the immune response to mycobacterial infection. OBJECTIVE: To identify whether iNOS expression and cytokine release of PBM are upregulated and have a connection in TB infection. DESIGN: The expression of iNOS immunoreactivity on PBM from TB patients and normal subjects was measured by loading with anti-macrophage iNOS polyclonal primary antibody analyzed by flow cytometry. Expression of iNOS mRNA in PBM was detected by RT-PCR. The spontaneous generation of nitrite and cytokines (IL-1 beta and TNF-alpha) by cultured monocytes was also determined. RESULTS: Compared to normal subjects, iNOS immuno-reactivity, the capacity for spontaneous nitrite generation and the level of TNF-alpha or IL-1 beta secretion of PBM were significantly higher in TB patients. The amount of nitrite, TNF-alpha and IL-1 beta released from PBM of TB patients was inhibited by NG-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of NOS. The level of iNOS immunoreactivity on PBM was highly correlated with nitrite generation both in all the subjects studied and in TB patients alone. Spontaneous TNF-alpha production showed a stronger correlation with nitrite production than with IL-1 beta. CONCLUSION: The NO and cytokine synthase activities of monocytes appear to be concomitantly upregulated in response to mycobacterial infection. The enhanced NO generation by monocytes in TB patients may play an autoregulatory role in amplifying the synthesis of pro-inflammatory cytokines.