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1.
J Allergy Clin Immunol Pract ; 7(6): 1970-1985.e4, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30877075

RESUMO

BACKGROUND: Although autoimmunity and hyperinflammation secondary to recombination activating gene (RAG) deficiency have been associated with delayed diagnosis and even death, our current understanding is limited primarily to small case series. OBJECTIVE: Understand the frequency, severity, and treatment responsiveness of autoimmunity and hyperinflammation in RAG deficiency. METHODS: In reviewing the literature and our own database, we identified 85 patients with RAG deficiency, reported between 2001 and 2016, and compiled the largest case series to date of 63 patients with prominent autoimmune and/or hyperinflammatory pathology. RESULTS: Diagnosis of RAG deficiency was delayed a median of 5 years from the first clinical signs of immune dysregulation. Most patients (55.6%) presented with more than 1 autoimmune or hyperinflammatory complication, with the most common etiologies being cytopenias (84.1%), granulomas (23.8%), and inflammatory skin disorders (19.0%). Infections, including live viral vaccinations, closely preceded the onset of autoimmunity in 28.6% of cases. Autoimmune cytopenias had early onset (median, 1.9, 2.1, and 2.6 years for autoimmune hemolytic anemia, immune thrombocytopenia, and autoimmune neutropenia, respectively) and were refractory to intravenous immunoglobulin, steroids, and rituximab in most cases (64.7%, 73.7%, and 71.4% for autoimmune hemolytic anemia, immune thrombocytopenia, and autoimmune neutropenia, respectively). Evans syndrome specifically was associated with lack of response to first-line therapy. Treatment-refractory autoimmunity/hyperinflammation prompted hematopoietic stem cell transplantation in 20 patients. CONCLUSIONS: Autoimmunity/hyperinflammation can be a presenting sign of RAG deficiency and should prompt further evaluation. Multilineage cytopenias are often refractory to immunosuppressive treatment and may require hematopoietic cell transplantation for definitive management.

2.
Contemp Clin Trials ; 78: 27-33, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30659924

RESUMO

Asthma causes enormous suffering and cost for children in the US and around the world [1-3]. Co-morbid gastroesophageal reflux disease (GERD) makes asthma management more difficult due to increased symptoms. Proton pump inhibitor (PPI) drugs are effective at improving to GERD symptoms, however they have demonstrated only modest and variable effects on asthma control in the setting of co-morbid GERD. Importantly, PPI metabolism and efficacy depend on CYP2C19 genotype. The Genotype Tailored Treatment of Symptomatic Acid Reflux in Children with Uncontrolled Asthma (GenARA) study is a randomized, double-blind, placebo-controlled trial to determine if genotype-tailored PPI dosing improves asthma symptoms among children with inadequately controlled asthma and GERD symptoms. This study has an innovative design to both assess the efficacy of genotype-tailored PPI dosing and perform pharmacokinetic modeling of the oral PPI Lansoprazole. Children ages 6-17 years old with clinician-diagnosed asthma and mild GERD symptoms will submit a saliva sample for CYP2C19 genotyping. Participants will undergo a two-step randomization to: (1) genotype-tailored versus conventional dosing of open-label oral lansoprazole for pharmacokinetic modeling, and (2) genotype-tailored lansoprazole daily versus placebo for 24 weeks to determine the effect of genotype-tailored PPI dosing on asthma control. Measures of asthma control, spirometry, and nasal washes during acute illnesses will be collected at 8-week intervals throughout the study. GenARA will better define the effects of CYP2C19 genotype on the dose response of lansoprazole in children and adolescents and assess if a novel dosing regimen improves GERD and asthma control.

3.
Chem Commun (Camb) ; 54(78): 11053, 2018 09 27.
Artigo em Inglês | MEDLINE | ID: mdl-30225473

RESUMO

Correction for 'Determination of ferric ions using surface-enhanced Raman scattering based on desferrioxamine-functionalized silver nanoparticles' by Fei Yan et al., Chem. Commun., 2013, 49, 7962-7964.

4.
J Immunol ; 194(12): 6123-32, 2015 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-25957169

RESUMO

Mycoplasma pneumoniae is an extracellular pathogen that colonizes mucosal surfaces of the respiratory tract and is associated with asthma exacerbations. Previous reports demonstrate that surfactant protein-A (SP-A) binds live M. pneumoniae and mycoplasma membrane fractions (MMF) with high affinity. Humans express a repertoire of single-amino acid genetic variants of SP-A that may be associated with lung disease, and our findings demonstrate that allelic differences in SP-A2 (Gln223Lys) affect the binding to MMF. We show that SP-A(-/-) mice are more susceptible to MMF exposure and have significant increases in mucin production and neutrophil recruitment. Novel humanized SP-A2-transgenic mice harboring the hSP-A2 223K allele exhibit reduced neutrophil influx and mucin production in the lungs when challenged with MMF compared with SP-A(-/-) mice. Conversely, mice expressing hSP-A2 223Q have increased neutrophil influx and mucin production that are similar to SP-A(-/-) mice. Using tracheal epithelial cell cultures, we show that enhanced mucin production to MMF occurs in the absence of SP-A and is not dependent upon neutrophil recruitment. Increased phosphorylation of the epidermal growth factor receptor (EGFR) was evident in the lungs of MMF-challenged mice when SP-A was absent. Pharmacologic inhibition of EGFR prior to MMF challenge dramatically reduced mucin production in SP-A(-/-) mice. These findings suggest a protective role for SP-A in limiting MMF-stimulated mucin production that occurs through interference with EGFR-mediated signaling. SP-A interaction with the EGFR signaling pathway appears to occur in an allele-specific manner that may have important implications for SP-A polymorphisms in human diseases.


Assuntos
Variação Genética , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Mycoplasma pneumoniae/imunologia , Mycoplasma pneumoniae/metabolismo , Pneumonia por Mycoplasma/genética , Pneumonia por Mycoplasma/imunologia , Proteína A Associada a Surfactante Pulmonar/genética , Animais , Membrana Celular/imunologia , Membrana Celular/metabolismo , Quimiotaxia/imunologia , Receptores ErbB/metabolismo , Feminino , Expressão Gênica , Ordem dos Genes , Marcação de Genes , Vetores Genéticos/genética , Humanos , Pulmão/imunologia , Pulmão/metabolismo , Pulmão/microbiologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Mucina-5AC/genética , Mucina-5AC/metabolismo , Muco/metabolismo , Neutrófilos/imunologia , Neutrófilos/metabolismo , Fosforilação , Pneumonia por Mycoplasma/metabolismo , Ligação Proteica , Proteína A Associada a Surfactante Pulmonar/metabolismo
5.
Artigo em Inglês | MEDLINE | ID: mdl-25673984

RESUMO

Chronic obstructive pulmonary disease (COPD) is one of the most common chronic illnesses in the world. The disease encompasses emphysema, chronic bronchitis, and small airway obstruction and can be caused by environmental exposures, primarily cigarette smoking. Since only a small subset of smokers develop COPD, it is believed that host factors interact with the environment to increase the propensity to develop disease. The major pathogenic factors causing disease include infection and inflammation, protease and antiprotease imbalance, and oxidative stress overwhelming antioxidant defenses. In this review, we will discuss the major environmental and host sources for oxidative stress; discuss how oxidative stress regulates chronic bronchitis; review the latest information on genetic predisposition to COPD, specifically focusing on oxidant/antioxidant imbalance; and review future antioxidant therapeutic options for COPD. The complexity of COPD will necessitate a multi-target therapeutic approach. It is likely that antioxidant supplementation and dietary antioxidants will have a place in these future combination therapies.


Assuntos
Antioxidantes/metabolismo , Pulmão/metabolismo , Oxidantes/metabolismo , Estresse Oxidativo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Animais , Antioxidantes/uso terapêutico , Epigênese Genética , Regulação da Expressão Gênica , Interação Gene-Ambiente , Humanos , Pulmão/fisiopatologia , Mucinas/genética , Mucinas/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/etiologia , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Fatores de Risco , Fumar/efeitos adversos , Fumar/metabolismo
6.
Am J Respir Cell Mol Biol ; 51(4): 568-74, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24816162

RESUMO

Diacetyl (DA), a component of artificial butter flavoring, has been linked to the development of bronchiolitis obliterans (BO), a disease of airway epithelial injury and airway fibrosis. The epidermal growth factor receptor ligand, amphiregulin (AREG), has been implicated in other types of epithelial injury and lung fibrosis. We investigated the effects of DA directly on the pulmonary epithelium, and we hypothesized that DA exposure would result in epithelial cell shedding of AREG. Consistent with this hypothesis, we demonstrate that DA increases AREG by the pulmonary epithelial cell line NCI-H292 and by multiple independent primary human airway epithelial donors grown under physiologically relevant conditions at the air-liquid interface. Furthermore, we demonstrate that AREG shedding occurs through a TNF-α-converting enzyme (TACE)-dependent mechanism via inhibition of TACE activity in epithelial cells using the small molecule inhibitor, TNF-α protease inhibitor-1, as well as TACE-specific small inhibitor RNA. Finally, we demonstrate supportive in vivo results showing increased AREG transcript and protein levels in the lungs of rodents with DA-induced BO. In summary, our novel in vitro and in vivo observations suggest that further study of AREG is warranted in the pathogenesis of DA-induced BO.


Assuntos
Bronquiolite Obliterante/induzido quimicamente , Diacetil/toxicidade , Família de Proteínas EGF/metabolismo , Células Epiteliais/efeitos dos fármacos , Aromatizantes/toxicidade , Mucosa Respiratória/efeitos dos fármacos , Proteínas ADAM/antagonistas & inibidores , Proteínas ADAM/genética , Proteínas ADAM/metabolismo , Proteína ADAM17 , Anfirregulina , Bronquiolite Obliterante/genética , Bronquiolite Obliterante/metabolismo , Linhagem Celular , Relação Dose-Resposta a Droga , Família de Proteínas EGF/genética , Inibidores Enzimáticos/farmacologia , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Humanos , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Interferência de RNA , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Fatores de Tempo , Transfecção , Regulação para Cima
7.
Am J Respir Cell Mol Biol ; 50(4): 684-9, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24325600

RESUMO

Neutrophil elastase (NE) is a major inflammatory mediator in cystic fibrosis (CF) that is a robust predictor of lung disease progression. NE directly causes airway injury via protease activity, and propagates persistent neutrophilic inflammation by up-regulation of neutrophil chemokine expression. Despite its key role in the pathogenesis of CF lung disease, there are currently no effective antiprotease therapies available to patients with CF. Although heparin is an effective antiprotease and anti-inflammatory agent, its anticoagulant activity prohibits its use in CF, due to risk of pulmonary hemorrhage. In this report, we demonstrate the efficacy of a 2-O, 3-O-desulfated heparin (ODSH), a modified heparin with minimal anticoagulant activity, to inhibit NE activity and to block NE-induced airway inflammation. Using an established murine model of intratracheal NE-induced airway inflammation, we tested the efficacy of intratracheal ODSH to block NE-generated neutrophil chemoattractants and NE-triggered airway neutrophilic inflammation. ODSH inhibited NE-induced keratinocyte-derived chemoattractant and high-mobility group box 1 release in bronchoalveolar lavage. ODSH also blocked NE-stimulated high-mobility group box 1 release from murine macrophages in vitro, and inhibited NE activity in functional assays consistent with prior reports of antiprotease activity. In summary, this report suggests that ODSH is a promising antiprotease and anti-inflammatory agent that may be useful as an airway therapy in CF.


Assuntos
Anti-Inflamatórios/farmacologia , Proteína HMGB1/metabolismo , Heparina/análogos & derivados , Elastase de Leucócito/antagonistas & inibidores , Pulmão/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Pneumonia/tratamento farmacológico , Inibidores de Proteases/farmacologia , Animais , Linhagem Celular , Quimiotaxia/efeitos dos fármacos , Modelos Animais de Doenças , Heparina/farmacologia , Interleucina-13 , Elastase de Leucócito/metabolismo , Pulmão/metabolismo , Pulmão/fisiopatologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Infiltração de Neutrófilos/efeitos dos fármacos , Pneumonia/induzido quimicamente , Pneumonia/metabolismo , Pneumonia/fisiopatologia
8.
Artigo em Inglês | MEDLINE | ID: mdl-23378755

RESUMO

There is evidence that proteases and antiproteases participate in the iron homeostasis of cells and living systems. We tested the postulate that α-1 antitrypsin (A1AT) polymorphism and the consequent deficiency of this antiprotease in humans are associated with a systemic disruption in iron homeostasis. Archived plasma samples from Alpha-1 Foundation (30 MM, 30 MZ, and 30 ZZ individuals) were analyzed for A1AT, ferritin, transferrin, and C-reactive protein (CRP). Plasma samples were also assayed for metals using inductively coupled plasma atomic emission spectroscopy (ICPAES). Plasma levels of A1AT in MZ and ZZ individuals were approximately 60% and 20% of those for MM individuals respectively. Plasma ferritin concentrations in those with the ZZ genotype were greater relative to those individuals with either MM or MZ genotype. Plasma transferrin for MM, MZ, and ZZ genotypes showed no significant differences. Linear regression analysis revealed a significant (negative) relationship between plasma concentrations of A1AT and ferritin while that between A1AT and transferrin levels was not significant. Plasma CRP concentrations were not significantly different between MM, MZ, and ZZ individuals. ICPAES measurement of metals confirmed elevated plasma concentrations of nonheme iron among ZZ individuals. Nonheme iron concentrations correlated (negatively) with levels of A1AT. A1AT deficiency is associated with evidence of a disruption in iron homeostasis with plasma ferritin and nonheme iron concentrations being elevated among those with the ZZ genotype.


Assuntos
Ferritinas/sangue , Transferrina/análise , Deficiência de alfa 1-Antitripsina/genética , alfa 1-Antitripsina/sangue , Alumínio/sangue , Proteína C-Reativa/análise , Cobre/sangue , Feminino , Genótipo , Homeostase , Humanos , Ferro/sangue , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Polimorfismo Genético , Espectrofotometria Atômica
9.
Am J Physiol Lung Cell Mol Physiol ; 304(6): L394-400, 2013 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-23316069

RESUMO

Cystic Fibrosis (CF) is a chronic lung disease characterized by chronic neutrophilic airway inflammation and increased levels of neutrophil elastase (NE) in the airways. We have previously reported that NE treatment triggers cell cycle arrest. Cell cycle arrest can lead to senescence, a complete loss of replicative capacity. Importantly, senescent cells can be proinflammatory and would perpetuate CF chronic inflammation. By immunohistochemistry, we evaluated whether airway sections from CF and control subjects expressed markers of senescence, including p16(INK4a) (p16), a cyclin-dependent kinase inhibitor, phospho-Histone H2A.X (γH2A.X), and phospho-checkpoint 2 kinase (phospho-Chk2), which are also DNA damage response markers. Compared with airway epithelium from control subjects, CF airway epithelium had increased levels of expression of all three senescence markers. We hypothesized that the high load of NE in the CF airway triggers epithelial senescence by upregulating expression of p16, which inhibits cyclin-dependent kinase 4 (CDK4). Normal human bronchial epithelial (NHBE) cells, cultured in air-liquid interface were treated with NE (0, 200, and 500 nM) to induce visible injury. Total cell lysates were collected and evaluated by Western analysis for p16 protein expression and CDK4 kinase activity. NE significantly increased p16 expression and decreased CDK4 kinase activity in NHBE cells. These results support the concept that NE triggers expression of senescence markers in CF airway epithelial cells.


Assuntos
Senescência Celular , Fibrose Cística/metabolismo , Mucosa Respiratória/metabolismo , Biomarcadores/metabolismo , Brônquios/patologia , Estudos de Casos e Controles , Células Cultivadas , Quinase do Ponto de Checagem 2 , Quinase 4 Dependente de Ciclina/metabolismo , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Fibrose Cística/genética , Fibrose Cística/patologia , Dano ao DNA , Histonas/metabolismo , Humanos , Elastase de Leucócito/fisiologia , Proteínas Serina-Treonina Quinases/metabolismo , Mucosa Respiratória/fisiopatologia , Telômero/genética , Telômero/metabolismo , Regulação para Cima
10.
J Biol Chem ; 288(7): 4681-91, 2013 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-23275341

RESUMO

NADPH:quinone oxidoreductase 1 (NQO1) is recognized as a major susceptibility gene for ozone-induced pulmonary toxicity. In the absence of NQO1 as can occur by genetic mutation, the human airway is protected from harmful effects of ozone. We recently reported that NQO1-null mice are protected from airway hyperresponsiveness and pulmonary inflammation following ozone exposure. However, NQO1 regenerates intracellular antioxidants and therefore should protect the individual from oxidative stress. To explain this paradox, we tested whether in the absence of NQO1 ozone exposure results in increased generation of A(2)-isoprostane, a cyclopentenone isoprostane that blunts inflammation. Using GC-MS, we found that NQO1-null mice had greater lung tissue levels of D(2)- and E(2)-isoprostanes, the precursors of J(2)- and A(2)-isoprostanes, both at base line and following ozone exposure compared with congenic wild-type mice. We confirmed in primary cultures of normal human bronchial epithelial cells that A(2)-isoprostane inhibited ozone-induced NF-κB activation and IL-8 regulation. Furthermore, we determined that A(2)-isoprostane covalently modified the active Cys(179) domain in inhibitory κB kinase in the presence of ozone in vitro, thus establishing the biochemical basis for A(2)-isoprostane inhibition of NF-κB. Our results demonstrate that host factors may regulate pulmonary susceptibility to ozone by regulating the generation of A(2)-isoprostanes in the lung. These observations provide the biochemical basis for the epidemiologic observation that NQO1 regulates pulmonary susceptibility to ozone.


Assuntos
Isoprostanos/química , NAD(P)H Desidrogenase (Quinona)/fisiologia , Ozônio/química , Animais , Linhagem Celular , Cisteína/genética , Humanos , Inflamação , Interleucina-8/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Espectrometria de Massas/métodos , Camundongos , Camundongos Endogâmicos C57BL , Modelos Biológicos , NAD(P)H Desidrogenase (Quinona)/metabolismo , NF-kappa B/metabolismo , Oxirredução
11.
Am J Physiol Lung Cell Mol Physiol ; 303(3): L181-8, 2012 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-22659878

RESUMO

Mucous cell metaplasia (MCM) and neutrophil-predominant airway inflammation are pathological features of chronic inflammatory airway diseases. A signature feature of MCM is increased expression of a major respiratory tract mucin, MUC5AC. Neutrophil elastase (NE) upregulates MUC5AC in primary airway epithelial cells by generating reactive oxygen species, and this response is due in part to upregulation of NADPH quinone oxidoreductase 1 (NQO1) activity. Delivery of NE directly to the airway triggers inflammation and MCM and increases synthesis and secretion of MUC5AC protein from airway epithelial cells. We hypothesized that NE-induced MCM is mediated in vivo by NQO1. Male wild-type and Nqo1-null mice (C57BL/6 background) were exposed to human NE (50 µg) or vehicle via oropharyngeal aspiration on days 1, 4, and 7. On days 8 and 11, lung tissues and bronchoalveolar lavage (BAL) samples were obtained and evaluated for MCM, inflammation, and oxidative stress. MCM, inflammation, and production of specific cytokines, granulocyte-macrophage colony-stimulating factor, macrophage inflammatory protein-2, interleukin-4, and interleukin-5 were diminished in NE-treated Nqo1-null mice compared with NE-treated wild-type mice. However, in contrast to the role of NQO1 in vitro, we demonstrate that NE-treated Nqo1-null mice had greater levels of BAL and lung tissue lipid carbonyls and greater BAL iron on day 11, all consistent with increased oxidative stress. NQO1 is required for NE-induced inflammation and MCM. This model system demonstrates that NE-induced MCM directly correlates with inflammation, but not with oxidative stress.


Assuntos
Inflamação/etiologia , Elastase de Leucócito/metabolismo , Metaplasia/etiologia , Metaplasia/patologia , NAD(P)H Desidrogenase (Quinona)/fisiologia , Estresse Oxidativo , Mucosa Respiratória/patologia , Animais , Lavagem Broncoalveolar , Células Cultivadas , Citocinas/metabolismo , Humanos , Técnicas Imunoenzimáticas , Inflamação/metabolismo , Inflamação/patologia , Ferro/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Masculino , Metaplasia/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mucina-5AC/metabolismo , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Mucosa Respiratória/metabolismo
12.
Br J Pharmacol ; 165(7): 2365-77, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22013997

RESUMO

BACKGROUND AND PURPOSE: Inhalation of a ß-adrenoceptor agonist (ß-agonist) is first-line asthma therapy, used for both prophylaxis against, and acute relief of, bronchoconstriction. However, repeated clinical use of ß-agonists leads to impaired bronchoprotection and, in some cases, adverse patient outcomes. Mechanisms underlying this ß(2) -adrenoceptor dysfunction are not well understood, due largely to the lack of a comprehensive animal model and the uncertainty as to whether or not bronchorelaxation in mice is mediated by ß(2) -adrenoceptors. Thus, we aimed to develop a mouse model that demonstrated functional ß-agonist-induced ß(2) -adrenoceptor desensitization in the context of allergic inflammatory airway disease. EXPERIMENTAL APPROACH: We combined chronic allergen exposure with repeated ß-agonist inhalation in allergen-treated BALB/C mice and examined the contribution of ß(2) -adrenoceptors to albuterol-induced bronchoprotection using FVB/NJ mice with genetic deletion of ß(2) -adrenoceptors (KO). Associated inflammatory changes - cytokines (ELISA), cells in bronchoalevolar lavage and airway remodelling (histology) and ß(2) -adrenoceptor density (radioligand binding) - were also measured. KEY RESULTS ß(2) -Adrenoceptors mediated albuterol-induced bronchoprotection in mice. Chronic treatment with albuterol induced loss of bronchoprotection, associated with exacerbation of the inflammatory components of the asthma phenotype. CONCLUSIONS AND IMPLICATIONS: This animal model reproduced salient features of human asthma and linked loss of bronchoprotection with airway pathobiology. Accordingly, the model offers an advanced tool for understanding the mechanisms of the effects of chronic ß- agonist treatment on ß-adrenoceptor function in asthma. Such information may guide the clinical use of ß-agonists and provide insight into development of novel ß-adrenoceptor ligands for the treatment of asthma.


Assuntos
Agonistas de Receptores Adrenérgicos beta 2/administração & dosagem , Agonistas de Receptores Adrenérgicos beta 2/efeitos adversos , Pneumonia/etiologia , Administração por Inalação , Animais , Antiasmáticos/administração & dosagem , Antiasmáticos/efeitos adversos , Asma/complicações , Asma/tratamento farmacológico , Broncoconstrição/efeitos dos fármacos , Modelos Animais de Doenças , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Pneumonia/patologia , Pneumonia/fisiopatologia , Receptores Adrenérgicos beta 2/deficiência , Receptores Adrenérgicos beta 2/efeitos dos fármacos , Receptores Adrenérgicos beta 2/genética
13.
Artigo em Inglês | MEDLINE | ID: mdl-21857781

RESUMO

Patients with chronic obstructive pulmonary disease (COPD) exhibit dominant features of chronic bronchitis, emphysema, and/or asthma, with a common phenotype of airflow obstruction. COPD pulmonary physiology reflects the sum of pathological changes in COPD, which can occur in large central airways, small peripheral airways, and the lung parenchyma. Quantitative or high-resolution computed tomography is used as a surrogate measure for assessment of disease progression. Different biological or molecular markers have been reported that reflect the mechanistic or pathogenic triad of inflammation, proteases, and oxidants and correspond to the different aspects of COPD histopathology. Similar to the pathogenic triad markers, genetic variations or polymorphisms have also been linked to COPD-associated inflammation, protease-antiprotease imbalance, and oxidative stress. Furthermore, in recent years, there have been reports identifying aging-associated mechanistic markers as downstream consequences of the pathogenic triad in the lungs from COPD patients. For this review, the authors have limited their discussion to a review of mechanistic markers and genetic variations and their association with COPD histopathology and disease status.


Assuntos
Interação Gene-Ambiente , Inflamação , Estresse Oxidativo/genética , Inibidores de Proteases/metabolismo , Doença Pulmonar Obstrutiva Crônica , Envelhecimento/metabolismo , Envelhecimento/patologia , Biomarcadores , Estudo de Associação Genômica Ampla , Humanos , Imuno-Histoquímica , Inflamação/complicações , Inflamação/genética , Doença Pulmonar Obstrutiva Crônica/etiologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Espécies Reativas de Oxigênio/metabolismo , Testes de Função Respiratória , Sistema Respiratório/metabolismo , Sistema Respiratório/patologia , Sistema Respiratório/fisiopatologia , Fatores de Risco , Fumar/metabolismo , Fumar/fisiopatologia
14.
Am J Physiol Lung Cell Mol Physiol ; 299(3): L345-52, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20543006

RESUMO

In a mouse model of neutrophil elastase-induced bronchitis that exhibits goblet cell metaplasia and inflammation, we investigated the effects of intratracheal instillation of the MANS peptide, a peptide identical to the NH(2) terminus of the myristoylated alanine-rich C kinase substrate (MARCKS) on mucin protein airway secretion, inflammation, and airway reactivity. To induce mucus cell metaplasia in the airways, male BALB/c mice were treated repetitively with the serine protease, neutrophil elastase, on days 1, 4, and 7. On day 11, when goblet cell metaplasia was fully developed and profiles of proinflammatory cytokines were maximal, the animals were exposed to aerosolized methacholine after intratracheal instillation of MANS or a missense control peptide (RNS). MANS, but not RNS, attenuated the methacholine-stimulated secretion of the major respiratory mucin protein, Muc5ac (50% reduction). Concurrently, elastase-induced proinflammatory cytokines typically recovered in bronchoalveolar lavage (BAL), including KC, IL-1beta, IL-6, MCP-1, and TNFalpha, were reduced by the MANS peptide (mean levels decreased 50-60%). Secondary to the effects of MANS on mucin secretion and inflammation, mechanical lung function by forced oscillation technique was characterized with respect to airway reactivity in response to cumulative aerosol stimulation with serotonin. The MANS peptide was also found to effectively attenuate airway hyperresponsiveness to serotonin in this airway hypersecretory model. Collectively, these findings support the concept that even in airway epithelia remodeled with goblet cell metaplasia and in a state of mucin hypersecretion, exogenous attenuation of function of MARCKS protein via the MANS peptide decreases airway mucin secretion, inflammation, and hyperreactivity.


Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Proteínas de Membrana/antagonistas & inibidores , Mucina-5AC/metabolismo , Fragmentos de Peptídeos/farmacologia , Sistema Respiratório/efeitos dos fármacos , Sistema Respiratório/metabolismo , Aerossóis , Animais , Líquido da Lavagem Broncoalveolar/química , Citocinas/metabolismo , Células Caliciformes/patologia , Inflamação/prevenção & controle , Mediadores da Inflamação/metabolismo , Elastase de Leucócito/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Masculino , Metaplasia/induzido quimicamente , Cloreto de Metacolina/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Mucina-5AC/antagonistas & inibidores , Substrato Quinase C Rico em Alanina Miristoilada , Hipersensibilidade Respiratória/induzido quimicamente , Hipersensibilidade Respiratória/fisiopatologia , Serotonina
15.
Clin Transl Sci ; 2(5): 333-9, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20411049

RESUMO

Alpha-1-antitrypsin (A1AT) deficiency is characterized by increased neutrophil elastase (NE) activity and oxidative stress in the lung. We hypothesized that NE exposure generates reactive oxygen species by increasing lung non-heme iron. To test this hypothesis, we measured bronchoalveolar lavage (BAL) iron and ferritin levels, using inductively coupled plasma (ICP) optical emission spectroscopy and an ELISA respectively, in A1AT-deficient patients and healthy subjects. To confirm the role of NE in regulating lung iron homeostasis, we administered intratracheally NE or control buffer to rats and measured BAL and lung iron and ferritin. Our results demonstrated that A1AT-deficient patients and rats post-elastase exposure have elevated levels of iron and ferritin in the BAL. To investigate the mechanism of NE-induced increased iron levels, we exposed normal human airway epithelial cells to either NE or control vehicle in the presence or absence of ferritin, and quantified intracellular iron uptake using calcein fluorescence and ICP mass spectroscopy. We also tested whether NE degraded ferritin in vitro using ELISA and western analysis. We demonstrated in vitro that NE increased intracellular non-heme iron levels and degraded ferritin. Our results suggest that NE digests ferritin increasing the extracellular iron pool available for cellular uptake.


Assuntos
Brônquios/metabolismo , Ferro/química , Elastase de Leucócito/metabolismo , Deficiência de alfa 1-Antitripsina/metabolismo , Adulto , Animais , Feminino , Ferritinas/metabolismo , Fluoresceínas/química , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , Ratos , Fatores de Tempo
16.
Am J Respir Cell Mol Biol ; 41(1): 107-13, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19059883

RESUMO

One host susceptibility factor for ozone identified in epidemiologic studies is NAD(P)H quinone oxidoreductase 1 (NQO1). We hypothesized that after ozone exposure, NQO1 is required to increase 8-isoprostane (also known as F(2)-isoprostane) production, a recognized marker of ozone-induced oxidative stress, and to enhance airway inflammation and hyperresponsiveness. In this report, we demonstrate that in contrast to wild-type mice, NQO1-null mice are resistant to ozone and have blunted responses, including decreased production of F(2)-isoprostane and keratinocyte chemokine, decreased airway inflammation, and diminished airway hyperresponsiveness. Importantly, these results in mice correlate with in vitro findings in humans. In primary human airway epithelial cells, inhibition of NQO1 by dicumarol blocks ozone-induced F(2)-isoprostane production and IL-8 gene expression. Together, these results demonstrate that NQO1 modulates cellular redox status and influences the biologic and physiologic effects of ozone.


Assuntos
Brônquios/efeitos dos fármacos , Hiper-Reatividade Brônquica/enzimologia , Células Epiteliais/efeitos dos fármacos , NADPH Desidrogenase/metabolismo , Oxidantes/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Ozônio/toxicidade , Pneumonia/enzimologia , Animais , Brônquios/enzimologia , Brônquios/fisiopatologia , Hiper-Reatividade Brônquica/induzido quimicamente , Hiper-Reatividade Brônquica/fisiopatologia , Hiper-Reatividade Brônquica/prevenção & controle , Testes de Provocação Brônquica , Células Cultivadas , Quimiocinas/metabolismo , Dicumarol/farmacologia , Dinoprosta/análogos & derivados , Dinoprosta/metabolismo , Inibidores Enzimáticos/farmacologia , Células Epiteliais/enzimologia , Humanos , Interleucina-8/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , NAD(P)H Desidrogenase (Quinona) , NADPH Desidrogenase/antagonistas & inibidores , NADPH Desidrogenase/deficiência , NADPH Desidrogenase/genética , Oxirredução , Pneumonia/induzido quimicamente , Pneumonia/fisiopatologia , Pneumonia/prevenção & controle , Fatores de Tempo
17.
Am J Physiol Lung Cell Mol Physiol ; 295(6): L1018-27, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18849440

RESUMO

Migration of airway epithelial cells (AEC) is a necessary component of airway mucosal repair after injury. The cytokine IL-1beta, present in airway inflammation, has protean effects on constituent cells within the mucosa, but its effects on epithelial repair are not known. We examined migration in differentiated primary human AEC grown in air-liquid interface culture for up to 3 wk and in the 16HBE14o(-) cell line. Wounds were created by mechanical abrasion and followed to closure using digital microscopy. Concurrent treatment with IL-1beta (

Assuntos
Movimento Celular/efeitos dos fármacos , Células Epiteliais/metabolismo , Interleucina-1beta/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Mucosa Respiratória/metabolismo , Fator de Transcrição RelA/metabolismo , Asma/metabolismo , Linhagem Celular , Movimento Celular/fisiologia , Células Epiteliais/citologia , Proteínas de Choque Térmico HSP27/metabolismo , Humanos , Interleucina-1beta/metabolismo , MAP Quinase Quinase 4/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fosforilação/efeitos dos fármacos , Fosforilação/fisiologia , Mucosa Respiratória/citologia , Fator de Transcrição STAT3/metabolismo
18.
Int J Biochem Cell Biol ; 40(6-7): 1238-45, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18395488

RESUMO

Cystic fibrosis is the most common, inherited fatal disease in Caucasians. The major cause of morbidity and mortality is chronic lung disease due to infection and inflammation in the airways leading to bronchiectasis and respiratory failure. The signature pathologic features of CF lung disease including abnormal mucus obstructing airways, chronic infection with Staphylococcus aureus, Pseudomonas aeruginosa and other gram negative bacteria, and a robust neutrophil-dominant airway inflammation, are exacerbated by unopposed proteases present at high concentrations in the ASL. There is strong evidence that proteases, particularly neutrophil elastase, contribute to the pathology of CF by impairing mucociliary clearance, interfering with innate immune functions, and perpetuating neutrophilic inflammation. The mechanisms employed by proteases to impact airway function in CF will be reviewed.


Assuntos
Fibrose Cística/metabolismo , Peptídeo Hidrolases/metabolismo , Fibrose Cística/etiologia , Fibrose Cística/microbiologia , Fibrose Cística/fisiopatologia , Humanos , Inflamação/fisiopatologia , Elastase de Leucócito/metabolismo , Neutrófilos/enzimologia , Infecções por Pseudomonas/microbiologia , Infecções Estafilocócicas/microbiologia
19.
Am J Physiol Lung Cell Mol Physiol ; 293(3): L762-8, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17586698

RESUMO

Neutrophil elastase (NE), a serine protease present in high concentrations in the airways of cystic fibrosis patients, injures the airway epithelium. We examined the epithelial response to NE-mediated proteolytic injury. We have previously reported that NE treatment of airway epithelial cells causes a marked decrease in epithelial DNA synthesis and proliferation. We hypothesized that NE inhibits DNA synthesis by arresting cell cycle progression. Progression through the cell cycle is positively regulated by cyclin complexes and negatively regulated by cyclin-dependent kinase inhibitors (CKI). To test whether NE arrests cell cycle progression, we treated normal human bronchial epithelial (NHBE) cells with NE (50 nM) or control vehicle for 24 h and assessed the effect of treatment on the cell cycle by flow cytometry. NE treatment resulted in G(1) arrest. Arrest in G(1) phase may be the result of CKI inhibition of the cyclin E complex; therefore, we evaluated whether NE upregulated CKI expression and/or affected the interaction of CKIs with the cyclin E complex. Following NE or control vehicle treatment, expression of p27(Kip1), a member of the Cip/Kip family, was evaluated. NE increased p27(Kip1) gene and protein expression. NE increased the coimmunoprecipitation of p27(Kip1) with cyclin E complex, suggesting that p27(Kip1) inhibited cyclin E complex activity. Our results demonstrate that p27 is regulated by NE and is critical for NE-induced cell cycle arrest.


Assuntos
Ciclo Celular/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Elastase de Leucócito/farmacologia , Sistema Respiratório/citologia , Sistema Respiratório/efeitos dos fármacos , Células Cultivadas , Inibidor de Quinase Dependente de Ciclina p27/genética , Células Epiteliais/metabolismo , Fase G1/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Imunoprecipitação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sistema Respiratório/metabolismo , Fase S/efeitos dos fármacos
20.
Free Radic Biol Med ; 42(9): 1398-408, 2007 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-17395013

RESUMO

Neutrophil elastase (NE), a potent neutrophil inflammatory mediator, increases MUC5AC mucin gene expression through undefined pathways involving reactive oxygen species. To determine the source of NE-generated reactive oxygen species, we used pharmacologic inhibitors of oxidoreductases to test whether they blocked NE-regulated MUC5AC mRNA expression. We found that dicumarol, an inhibitor of the NADP(H):quinone oxidoreductase 1 (NQO1), inhibited MUC5AC mRNA expression in A549 lung adenocarcinoma cells and primary normal human bronchial epithelial cells. We further tested the role of NQO1 in mediating NE-induced MUC5AC expression by inhibiting NQO1 expression using short interfering RNA (siRNA). Transfection with siRNA specific for NQO1 suppressed NQO1 expression and significantly abrogated MUC5AC mRNA expression. NE treatment caused lipid peroxidation in A549 cells; this effect was inhibited by pretreatment with dicumarol, suggesting that NQO1 also regulates oxidant stress in A549 cells after NE exposure. NE exposure increased NQO1 protein and activity levels; NQO1 expression and activity were limited to the cytosol and did not translocate to the plasma membrane. Our results indicate that NQO1 has an important role as a key mediator of NE-regulated oxidant stress and MUC5AC mucin gene expression.


Assuntos
Regulação Neoplásica da Expressão Gênica/fisiologia , Elastase de Leucócito/metabolismo , Mucinas/genética , NAD(P)H Desidrogenase (Quinona)/metabolismo , Quinona Redutases/metabolismo , Adenocarcinoma , Linhagem Celular Tumoral , Humanos , Neoplasias Pulmonares , Mucina-5AC , NAD(P)H Desidrogenase (Quinona)/imunologia , RNA Mensageiro/genética , Transcrição Genética
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