RESUMO
The prevalence of asthma continues to increase and its optimal treatment remains a challenge. Here, we investigated the actions of lipoxin A(4) (LXA(4)) and its leukocyte receptor in pulmonary inflammation using a murine model of asthma. Allergen challenge initiated airway biosynthesis of LXA(4) and increased expression of its receptor. Administration of a stable analog of LXA(4) blocked both airway hyper-responsiveness and pulmonary inflammation, as shown by decreased leukocytes and mediators, including interleukin-5, interleukin-13, eotaxin, prostanoids and cysteinyl leukotrienes. Moreover, transgenic expression of human LXA(4) receptors in murine leukocytes led to significant inhibition of pulmonary inflammation and eicosanoid-initiated eosinophil tissue infiltration. Inhibition of airway hyper-responsiveness and allergic airway inflammation with a stable LXA(4) analog highlights a unique counter-regulatory profile for the LXA(4) system and its leukocyte receptor in airway responses. Moreover, our findings suggest that lipoxin and related pathways offer novel multi-pronged therapeutic approaches for human asthma.
Assuntos
Asma/tratamento farmacológico , Hiper-Reatividade Brônquica/tratamento farmacológico , Bronquite/tratamento farmacológico , Ácidos Hidroxieicosatetraenoicos/farmacologia , Lipoxinas , Receptores de Formil Peptídeo , Receptores de Lipoxinas , Animais , Asma/diagnóstico , Asma/metabolismo , Asma/fisiopatologia , Quimiocina CCL11 , Quimiocinas CC/metabolismo , Eosinófilos/efeitos dos fármacos , Eosinófilos/metabolismo , Eosinófilos/patologia , Interleucina-13/metabolismo , Interleucina-5/metabolismo , Leucotrienos/metabolismo , Masculino , Cloreto de Metacolina , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Prostaglandinas/metabolismo , Receptores de Superfície Celular/efeitos dos fármacos , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismoRESUMO
The cytokine IL-6 acts via a specific receptor complex that consists of the membrane-bound IL-6 receptor (mIL-6R) or the soluble IL-6 receptor (sIL-6R) and glycoprotein 130 (gp130). In this study, we investigated the role of IL-6R components in asthma. We observed increased levels of sIL-6R in the airways of patients with allergic asthma as compared to those in controls. In addition, local blockade of the sIL-6R in a murine model of late-phase asthma after OVA sensitization by gp130-fraction constant led to suppression of Th2 cells in the lung. By contrast, blockade of mIL-6R induced local expansion of Foxp3-positive CD4+CD25+ Tregs with increased immunosuppressive capacities. CD4+CD25+ but not CD4+CD25- lung T cells selectively expressed the IL-6R alpha chain and showed IL-6-dependent STAT-3 phosphorylation. Finally, in an in vivo transfer model of asthma in immunodeficient Rag1 mice, CD4+CD25+ T cells isolated from anti-IL-6R antibody-treated mice exhibited marked immunosuppressive and antiinflammatory functions. IL-6 signaling therefore controls the balance between effector cells and Tregs in the lung by means of different receptor components. Furthermore, inhibition of IL-6 signaling emerges as a novel molecular approach for the treatment of allergic asthma.
Assuntos
Asma/imunologia , Hipersensibilidade/imunologia , Pulmão/imunologia , Receptores de Interleucina-2/imunologia , Receptores de Interleucina-6/imunologia , Células Th2/imunologia , Adulto , Animais , Anticorpos/administração & dosagem , Anticorpos/imunologia , Asma/patologia , Proteínas de Ligação a DNA/imunologia , Feminino , Fatores de Transcrição Forkhead , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/imunologia , Humanos , Hipersensibilidade/patologia , Inflamação/imunologia , Inflamação/patologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Knockout , Ovalbumina/metabolismo , Receptores de Citocinas/imunologia , Fator de Transcrição STAT3 , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Células Th2/patologia , Transativadores/imunologiaRESUMO
The regulatory effect of substance P on respiration is mediated via neurokinin (NK) receptors. While previous studies suggest that NK-1 receptors are involved, little is known about the role NK-2 receptors in ventilatory responses to hypoxia. Ventilatory responses to acute hypoxia (8% O2 in N2) were measured by indirect plethysmography in unanaesthetized, unrestrained NK-1 receptor gene deficient (NK-1-/-) and wild-type mice. In additional experiments mice were treated with an NK-2 receptor antagonist prior to hypoxic challenge. Resting ventilatory parameters were not different between groups. NK-1-/- mice displayed significantly greater shortening of expiratory time and higher increase of breathing frequency during hypoxia than wild-type mice. Treatment with the NK-2 receptor antagonist SR 48968 (1 mg/kg) resulted in a further shortening of inspiratory and expiratory time in NK-1-/- but not wild-type mice. These results demonstrate that both NK-1 and NK-2 receptors are involved in the modification of ventilation in response to acute hypoxia.
Assuntos
Hipóxia/fisiopatologia , Receptores da Neurocinina-1/deficiência , Respiração/genética , Animais , Benzamidas/administração & dosagem , Feminino , Hipóxia/prevenção & controle , Masculino , Camundongos , Camundongos Knockout , Piperidinas/administração & dosagem , Pletismografia/métodos , Receptores da Neurocinina-2/antagonistas & inibidores , Respiração/efeitos dos fármacosRESUMO
Acute respiratory distress syndrome (ARDS) is a life-threatening ailment characterized by severe lung injury involving inflammatory cell recruitment to the lung, cytokine production, surfactant dysfunction, and up-regulation of nitric oxide synthase 2 (NOS2) resulting in nitric oxide (NO) production. We hypothesized that NO production from NOS2 expressed in lung parenchymal cells in a murine model of ARDS would correlate with abnormal surfactant function and reduced surfactant protein-B (SP-B) expression. Pulmonary responses to nebulized endotoxin (lipopolysaccharide, LPS) were evaluated in wild-type (WT) mice, NOS2 null (-/-) mice, and NOS2-chimeric animals derived from bone marrow transplantation. NOS2-/- animals exhibited significantly less physiologic lung dysfunction and loss of SP-B expression than did WT animals. However, lung neutrophil recruitment and bronchoalveolar lavage cytokine levels did not significantly differ between NOS2-/- and WT animals. Chimeric animals for NOS2 exhibited the phenotype of the recipient and therefore demonstrated that parenchymal production of NOS2 is critical for the development of LPS-induced lung injury. Furthermore, administration of NO donors, independent of cytokine stimulation, decreased SP-B promoter activity and mRNA expression in mouse lung epithelial cells. This study demonstrates that expression of NOS2 in lung epithelial cells is critical for the development of lung injury and mediates surfactant dysfunction independent of NOS2 inflammatory cell expression and cytokine production.
Assuntos
Lipopolissacarídeos/toxicidade , Pulmão/patologia , Óxido Nítrico Sintase/fisiologia , Proteína B Associada a Surfactante Pulmonar/biossíntese , Síndrome do Desconforto Respiratório/enzimologia , Aerossóis , Animais , Transplante de Medula Óssea , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Inflamação , Interleucina-6/análise , Contagem de Leucócitos , Lipopolissacarídeos/administração & dosagem , Pulmão/efeitos dos fármacos , Pulmão/enzimologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Animais , Óxido Nítrico/fisiologia , Óxido Nítrico Sintase/deficiência , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo II , Proteína A Associada a Surfactante Pulmonar/farmacologia , Proteína B Associada a Surfactante Pulmonar/genética , Proteína B Associada a Surfactante Pulmonar/fisiologia , Proteína C Associada a Surfactante Pulmonar/farmacologia , RNA Mensageiro/biossíntese , Quimera por Radiação , Síndrome do Desconforto Respiratório/induzido quimicamente , Síndrome do Desconforto Respiratório/patologia , Organismos Livres de Patógenos Específicos , Tensão Superficial/efeitos dos fármacos , Transcrição Gênica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/análiseRESUMO
Epidemiological studies have implicated wood smoke as a risk factor for exacerbating asthma. However, comparisons of findings in animal models with those in humans are currently not possible, because detailed clinically relevant measurements of pulmonary function are not available in animal studies. Brown Norway rats were immunized with ovalbumin and exposed to either filtered air or wood smoke at 1 mg particulate matter/m(3) for 70 days and challenged with allergen during the last 4 days of exposure. Baseline values for dynamic lung compliance were lower while functional residual capacity was increased in rats exposed to wood smoke compared to rats exposed to filtered air. IFN-gamma levels were reduced and IL-4 levels increased in the bronchoalveolar lavage fluid and blood plasma, inflammatory lesions in the lungs were 21% greater, and airway mucous cells/mm basal lamina were non-significantly increased in rats exposed to wood smoke compared to controls. Collectively, these studies suggest that the pulmonary function was affected in rats by exposure to wood smoke and this decline was associated with only minor increases in inflammation of the lung. Therefore, this animal model may be useful to elucidate the mechanisms of the decline in pulmonary function caused by environmental pollutants when asthmatics are exposed to allergen.
Assuntos
Poluentes Atmosféricos/toxicidade , Poluição do Ar/efeitos adversos , Hipersensibilidade Respiratória/induzido quimicamente , Fumaça/efeitos adversos , Madeira , Poluentes Atmosféricos/análise , Poluição do Ar/análise , Alérgenos/farmacologia , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Exposição por Inalação , Interferon gama/análise , Interleucina-4/análise , Masculino , Ovalbumina/farmacologia , Ratos , Ratos Endogâmicos BN , Testes de Função Respiratória , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/patologia , Mucosa Respiratória/efeitos dos fármacos , Mucosa Respiratória/patologia , Fumaça/análiseRESUMO
Asthma is one of the foremost contributors to morbidity and mortality in industrialized countries. Our objective was to characterize the acute response to allergen and to identify potentially novel molecular targets for pharmacological intervention in asthma. We therefore designed a study to identify genes whose regulation was altered following ovalbumin (OVA) challenge in the presence and absence of treatment with glucocorticoids in BALB/c mice. RNA was isolated from lungs for gene profiling from 8-week-old sensitized mice, 3 and 18 hours post OVA challenge on days 1, 4, and 7 of aerosol challenge. Taqman (real time RT-PCR) analysis of marker genes indicative of Th2 (IL-4, IL-13), eosinophil (RANTES, eotaxin), Th1/macrophage (IFNgamma) and epithelial cell (MUC5AC) phenotypes were used to characterize responses to allergen challenge. Histological evaluation of lungs from additional challenged animals revealed inflammatory infiltrates on days 4 and 7, but not on day 1 post challenge. We postulate that expression of IL-4, IL-13 and other genes by OVA at day 1 probably reflects activation of resident cells, whereas the fivefold increase in the number of regulated genes at day 7 reflects the contribution of recruited cells. Of the regulated genes, only a subset was counter-regulated by dexamethasone treatment. Although regulated genes included genes in many protein families, herein we report regulation of two proteases whose role in response to OVA challenge has not been characterized. This model will be used to generate disease hypotheses for which may play an important role in initiating disease pathology in this model.
Assuntos
Asma/genética , Animais , Antígenos/administração & dosagem , Asma/etiologia , Asma/imunologia , Asma/patologia , Citocinas/genética , Modelos Animais de Doenças , Perfilação da Expressão Gênica , Inflamação/etiologia , Inflamação/genética , Inflamação/imunologia , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Pulmão/imunologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologiaRESUMO
Individuals with asthma have increased levels of nitric oxide in their exhaled air. To explore its role, we have developed a regulatable transgenic mouse capable of overexpressing inducible nitric oxide synthase in a lung-specific fashion. The CC10-rtTA-NOS-2 mouse contains two transgenes, a reverse tetracycline transactivator under the control of the Clara cell protein promoter and the mouse nitric oxide synthase-2 (NOS-2) coding region under control of a tetracycline operator. Addition of doxycycline to the drinking water of CC10-rtTA-NOS-2 mice causes an increase in nitric oxide synthase-2 that is largely confined to the airway epithelium. The fraction of expired nitric oxide increases over the first 24 h from approximately 10 parts per billion to a plateau of approximately 20 parts per billion. There were no obvious differences between CC10-rtTA-NOS-2 mice, with or without doxycycline, and wild-type mice in lung histology, bronchoalveolar protein, total cell count, or count differentials. However, airway resistance was lower in CC10-rtTA-NOS-2 mice with doxycycline than in CC10-rtTA-NOS-2 mice without doxycycline or wild-type mice with doxycycline. Moreover, doxycycline-treated CC10-rtTA-NOS-2 mice were hyporesponsive to methacholine compared with other groups. These data suggest that increased nitric oxide in the airways has no proinflammatory effects per se and may have beneficial effects on pulmonary function.
Assuntos
Resistência das Vias Respiratórias/genética , Resistência das Vias Respiratórias/fisiologia , Pulmão/enzimologia , Pulmão/fisiologia , Óxido Nítrico Sintase/antagonistas & inibidores , Animais , Asma/enzimologia , Asma/metabolismo , Northern Blotting , Western Blotting , Líquido da Lavagem Broncoalveolar/citologia , Broncodilatadores/farmacologia , DNA Complementar/biossíntese , DNA Complementar/genética , Doxiciclina/metabolismo , Imuno-Histoquímica , Cloreto de Metacolina/farmacologia , Camundongos , Camundongos Transgênicos , Óxido Nítrico/metabolismo , Óxido Nítrico/fisiologia , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Mecânica Respiratória/genética , Mecânica Respiratória/fisiologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tetraciclina/metabolismo , Transativadores/genética , Transativadores/metabolismo , Transgenes , Uteroglobina/genéticaRESUMO
Here, we have demonstrated potent inhibition of allergen-mediated pulmonary inflammation and airway hyper-responsiveness by a novel dual-pronged action of LX's. Moreover, these results indicate that LX's play pivotal and previously unappreciated roles in regulating allergy and pulmonary inflammation.
Assuntos
Aspirina/farmacologia , Brônquios/efeitos dos fármacos , Brônquios/metabolismo , Hiper-Reatividade Brônquica/metabolismo , Ácidos Hidroxieicosatetraenoicos/metabolismo , Ácidos Hidroxieicosatetraenoicos/farmacologia , Lipoxinas , Receptores de Formil Peptídeo , Receptores de Lipoxinas , Animais , Brônquios/patologia , Hiper-Reatividade Brônquica/tratamento farmacológico , Hiper-Reatividade Brônquica/patologia , Humanos , Ácidos Hidroxieicosatetraenoicos/administração & dosagem , Inflamação/metabolismo , Inflamação/patologia , Leucócitos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismoRESUMO
Ambient ozone has been linked to the worsening of symptoms of patients with obstructive diseases such as chronic obstructive pulmonary disease (COPD) and asthma. We investigated the role of cathepsin S on ozone-induced airway hyperresponsiveness (AHR) and inflammation, using the selective cathepsin S inhibitor, Compound A. Balb/c mice were exposed to ozone at a concentration of 3 ppm or air for 3 h, following administration by gavage of Compound A or vehicle. Bronchoalveolar lavage (BAL) was performed 3 h and 20-24 h following exposure, AHR was measured at 20-24 h only. Ozone exposure, compared to air exposure increased BAL cathepsin S levels, AHR and BAL inflammatory cells. Compound A (30 mg kg(-1) p.o.) dosing compared to vehicle dosing inhibited ozone-induced AHR (-logPC100 vehicle: -0.70+/-0.12, n=8 vs. cathepsin S inhibitor: -1.30+/-0.06, P<0.001, n=8) at 20-24 h and BAL neutrophilia at 3 h and 20-24 h (P<0.05, n=6). Ozone exposure increased levels of BAL cytokines IL-6, TNF-alpha and IFN-gamma. Compound A reduced IL-6 at 3 h and 20-24 h (P<0.05, n=5) and TNF-alpha, at 20-24 h (P<0.05, n=6). These data indicate an important role for cathepsin S in the regulation of ozone-induced AHR and neutrophil cell recruitment and suggest that cathepsin S may be a target in the treatment of oxidative stress-induced AHR and inflammation.
Assuntos
Hiper-Reatividade Brônquica/fisiopatologia , Catepsinas/metabolismo , Inflamação/fisiopatologia , Ozônio/toxicidade , Poluentes Atmosféricos/toxicidade , Animais , Hiper-Reatividade Brônquica/induzido quimicamente , Lavagem Broncoalveolar , Catepsinas/antagonistas & inibidores , Sistemas de Liberação de Medicamentos , Inflamação/induzido quimicamente , Interferon gama/efeitos dos fármacos , Interferon gama/metabolismo , Interleucina-6/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Infiltração de Neutrófilos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fatores de Tempo , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Th1/Tc1 inflammation and remodeling responses characterized by tissue atrophy and destruction frequently coexist in human diseases and disorders. However, the mechanisms that are used by Th1/Tc1 cytokines, like IFN-gamma, to induce these responses have not been defined. To elucidate the mechanism(s) of IFN-gamma-induced tissue remodeling and destruction, we characterized the pathway that lung-targeted, transgenic IFN-gamma uses to induce alveolar remodeling in a murine pulmonary emphysema modeling system. In these mice, transgenic IFN-gamma caused epithelial cell DNA injury and apoptosis detectable with TUNEL (Roche) and dual annexin V and propidium iodide staining. These responses were associated with death receptor and mitochondrial apoptosis pathway activation. Importantly, apoptosis inhibition with a caspase inhibitor (N-benzylcarboxy-Val-Ala-Asp-fluoromethyl-ketone) or a null mutation of caspase-3 blocked this DNA injury and apoptosis response and significantly ameliorated IFN-gamma-induced emphysema. These interventions also ameliorated IFN-gamma-induced inflammation and decreased pulmonary protease burden. Selective cathepsin S inhibition and a null mutation of cathepsin S also decreased IFN-gamma-induced DNA injury, apoptosis, emphysema, inflammation, and protease accumulation. These studies demonstrate that cathepsin S-dependent epithelial cell apoptosis is a critical event in the pathogenesis of IFN-gamma-induced alveolar remodeling and emphysema. They also link inflammation, protease/antiprotease alterations, and protease-dependent apoptosis in the pathogenesis of Th1/Tc1 cytokine-induced tissue remodeling and destructive responses.
Assuntos
Apoptose/imunologia , Catepsinas/fisiologia , Interferon gama/fisiologia , Alvéolos Pulmonares/imunologia , Enfisema Pulmonar/imunologia , Enfisema Pulmonar/patologia , Mucosa Respiratória/citologia , Mucosa Respiratória/imunologia , Animais , Apoptose/genética , Catepsinas/biossíntese , Catepsinas/deficiência , Catepsinas/genética , Dano ao DNA/imunologia , Modelos Animais de Doenças , Indução Enzimática/genética , Indução Enzimática/imunologia , Humanos , Inflamação/enzimologia , Inflamação/imunologia , Inflamação/patologia , Pulmão/enzimologia , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Especificidade de Órgãos/genética , Especificidade de Órgãos/imunologia , Alvéolos Pulmonares/enzimologia , Alvéolos Pulmonares/patologia , Enfisema Pulmonar/enzimologia , Enfisema Pulmonar/genética , Mucosa Respiratória/enzimologia , Transdução de Sinais/genética , Transdução de Sinais/imunologiaRESUMO
Allergic airway responses cause proliferation of epithelial cells and mucus cell metaplasia (MCM), and the resolution of MCM involves reduction of cell numbers. The role of inflammation and apoptosis on this process was investigated in P-selectin +/+ and -/- mice sensitized and challenged with OVA by analyzing the expression and the role of regulators of apoptosis in metaplastic mucus cells. No differences were observed in MCM at 5 days of allergen exposure between +/+ and -/- mice, despite reduced IL-13 levels in -/- mice. Although IL-4 levels were similar in both -/- and +/+ mice, IL-13 and IL-5 levels had decreased and IFN-gamma levels were increased earlier in -/- compared with +/+ mice. MCM levels were decreased 4-fold at 7 days of allergen exposure in -/- mice and at 15 days in +/+ mice. The percentage of Bax-expressing mucus cells increased significantly at 7 days in -/- mice and at 10 days in +/+ mice. The Bax-positive mucus cells exhibited caspase-specific cleavage of cytokeratin 18. IFN-gamma caused Bax expression in IL-13-induced MCM in microdissected airway cultures. MCM remained significantly elevated in Bax -/- mice following 15 days of allergen exposure compared with +/+ mice, while the number of eosinophils was reduced in both Bax +/+ and -/- mice at 15 days. Together, these data demonstrate that reduced IL-13 levels were sufficient to elicit maximum MCM, that IFN-gamma induces Bax in metaplastic mucus cells, and that Bax plays a critical role in the resolution of MCM, but not in the resolution of eosinophils.
Assuntos
Alérgenos/administração & dosagem , Interferon gama/uso terapêutico , Pulmão/imunologia , Pulmão/patologia , Muco/citologia , Muco/imunologia , Proteínas Proto-Oncogênicas c-bcl-2/fisiologia , Proteínas Proto-Oncogênicas/fisiologia , Animais , Brônquios/imunologia , Brônquios/metabolismo , Contagem de Células , Técnicas de Cultura , Dissecação , Eosinofilia/imunologia , Inflamação/genética , Inflamação/imunologia , Inflamação/prevenção & controle , Pulmão/metabolismo , Masculino , Metaplasia/genética , Metaplasia/imunologia , Metaplasia/patologia , Metaplasia/prevenção & controle , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Muco/metabolismo , Ovalbumina/administração & dosagem , Selectina-P/genética , Proteínas Proto-Oncogênicas/biossíntese , Hipersensibilidade Respiratória/genética , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/patologia , Hipersensibilidade Respiratória/prevenção & controle , Proteína X Associada a bcl-2RESUMO
Inflammatory responses induced by allergen exposure cause mucous cell metaplasia (MCM) by differentiation of existing and proliferating epithelial cells into mucus-storing cells. Airway epithelia have various mechanisms that resolve these changes to form normal airway epithelia. In this report, we first investigated the state of mucous cell metaplasia and the mechanisms by which MCM is reduced despite continued exposures to allergen. After 5 days of allergen exposure, extensive MCM had developed but was reduced when allergen challenge was continued for 15 days. During this exposure period, IL-13 levels decreased and IFN-gamma levels increased in the bronchoalveolar lavage fluid. In contrast, IL-13 levels decreased but IFN-gamma was not detected at any time point during the resolution of MCM following cessation of allergen exposure. Instillation of IFN-gamma but not anti-Fas caused accelerated resolution of MCM and MCM was not resolved in Stat1-deficient mice exposed to allergen for 15 days, confirming that IFN-gamma is crucial for reducing MCM during prolonged exposures to allergen. IFN-gamma but not anti-Fas induced apoptotic cell death in proliferating normal human bronchial epithelial cells and in human bronchial epithelial cells from subjects with asthma. The apoptotic effect of IFN-gamma was caspase dependent and was inhibited by IL-13, indicating that the Th2 milieu in asthmatics may maintain MCM by preventing cell death in metaplastic mucous cells. These studies could be useful in the understanding of deficiencies leading to chronicity in airway changes and designing novel therapies to reverse MCM and airway obstruction in asthmatics.
Assuntos
Apoptose , Asma/patologia , Interferon gama/farmacologia , Mucosa Respiratória/patologia , Receptor fas/farmacologia , Adulto , Alérgenos/imunologia , Animais , Brônquios/citologia , Células Cultivadas , Proteínas de Ligação a DNA/fisiologia , Humanos , Hipersensibilidade/imunologia , Hipersensibilidade/patologia , Interferon gama/antagonistas & inibidores , Interleucina-13/farmacologia , Cinética , Masculino , Metaplasia , Camundongos , Camundongos Endogâmicos C57BL , Ovalbumina/imunologia , Mucosa Respiratória/efeitos dos fármacos , Fator de Transcrição STAT1 , Transativadores/fisiologiaRESUMO
We postulated that the seleno-organic compound ebselen would attenuate neutrophil recruitment and activation after aerosolized challenge with endotoxin (LPS) through its effect as an antioxidant and inhibitor of gene activation. Rats were given ebselen (1-100 mg/kg i.p.) followed by aerosolized LPS exposure (0.3 mg/ml for 30 min). Airway inflammatory indices were measured 4 h postchallenge. Bronchoalveolar lavage (BAL) fluid cellularity and myeloperoxidase activity were used as a measure of neutrophil recruitment and activation. RT-PCR analysis was performed in lung tissue to assess gene expression of TNF-alpha, cytokine-induced neutrophil chemoattractant-1 (CINC-1), macrophage-inflammatory protein-2 (MIP-2), ICAM-1, IL-10, and inducible NO synthase. Protein levels in lung and BAL were also determined by ELISA. Ebselen pretreatment inhibited neutrophil influx and activation as assessed by BAL fluid cellularity and myeloperoxidase activity in cell-free BAL and BAL cell homogenates. This protective effect was accompanied by a significant reduction in lung and BAL fluid TNF-alpha and IL-1 beta protein and/or mRNA levels. Ebselen pretreatment also prevented lung ICAM-1 mRNA up-regulation in response to airway challenge with LPS. This was not a global effect of ebselen on LPS-induced gene expression, because the rise in lung and BAL CINC-1 and MIP-2 protein levels were unaffected as were lung mRNA expressions for CINC-1, MIP-2, IL-10, and inducible NO synthase. These data suggest that the anti-inflammatory properties of ebselen are achieved through an inhibition of lung ICAM-1 expression possibly through an inhibition of TNF-alpha and IL-1 beta, which are potent neutrophil recruiting mediators and effective inducers of ICAM-1 expression.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Azóis/administração & dosagem , Quimiocinas CXC/biossíntese , Peptídeos e Proteínas de Sinalização Intercelular , Lipopolissacarídeos/toxicidade , Pulmão/efeitos dos fármacos , Pulmão/patologia , Infiltração de Neutrófilos/efeitos dos fármacos , Infiltração de Neutrófilos/imunologia , Compostos Organosselênicos/administração & dosagem , Aerossóis , Animais , Azóis/uso terapêutico , Brônquios/efeitos dos fármacos , Brônquios/imunologia , Brônquios/metabolismo , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Quimiocina CXCL1 , Quimiocina CXCL2 , Quimiocinas CXC/fisiologia , Fatores Quimiotáticos/biossíntese , Modelos Animais de Doenças , Substâncias de Crescimento/biossíntese , Inflamação/imunologia , Injeções Intraperitoneais , Molécula 1 de Adesão Intercelular/biossíntese , Interleucina-1/antagonistas & inibidores , Interleucina-1/biossíntese , Interleucina-1/genética , Isoindóis , Pulmão/imunologia , Masculino , Monocinas/biossíntese , Compostos Organosselênicos/uso terapêutico , RNA Mensageiro/antagonistas & inibidores , RNA Mensageiro/biossíntese , Ratos , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/genética , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/imunologiaRESUMO
Human asthma is associated with airway infiltration by T helper 2 (TH2) lymphocytes. We observed reduced expression of the TH1 transcription factor, T-bet, in T cells from airways of patients with asthma compared with that in T cells from airways of nonasthmatic patients, suggesting that loss of T-bet might be associated with asthma. Mice with a targeted deletion of the T-bet gene and severe combined immunodeficient mice receiving CD4+ cells from T-bet knockout mice spontaneously demonstrated multiple physiological and inflammatory features characteristic of asthma. Thus, T-bet deficiency, in the absence of allergen exposure, induces a murine phenotype reminiscent of both acute and chronic human asthma.