RESUMO
INTRODUCTION: Pulmonary emphysema is characterized by destruction of alveoli leading to inadequate oxygenation, disability and frequently death. This destruction was understood so far as irreversible. Published data has shown that ATRA (All Trans Retinoic Acid) reverses elastase-induced emphysema in rats. However, the molecular mechanisms governing regeneration process are so far unknown. OBJECTIVE: To examine the therapeutic potential of ATRA on various molecular pathways and their coordination towards governance of alveolar epithelial regeneration in emphysematous rats. METHODS: Emphysema was induced by elastase versus saline in Sprague-Dawley rats. On days 26-37, rats received daily intraperitoneal injections with ATRA (500⯵g/kg b.w.) versus olive-oil. Lungs were removed at day 38 for histopathology and investigation of relative mRNA and protein expressions. RESULTS: Histopathological analysis has shown that losses of alveoli were recovered in therapy (EA) group. Moreover, expressions of markers genes for alveolar cell proliferation, differentiation and EMT events at mRNA and protein levels were significantly increased in EA group than emphysema group (ES). Upon validation at genomics level, expressions of components of Notch, Hedgehog, Wnt, BMP and TGFß pathways were significantly attenuated in EA group when compared with ES and were well comparable with the healthy group. CONCLUSION: Therapeutic supplementation of ATRA rectifies the deregulated Notch, Hedgehog, Wnt, BMP and TGFß pathways in emphysema condition, resulting in alveolar epithelium regeneration. Hence, ATRA may prove to be a potential drug in the treatment of emphysema. Nevertheless, elaborated studies are to be conducted.
Assuntos
Alvéolos Pulmonares/efeitos dos fármacos , Enfisema Pulmonar/tratamento farmacológico , Regeneração/efeitos dos fármacos , Tretinoína/uso terapêutico , Animais , Aquaporina 4/genética , Peso Corporal/efeitos dos fármacos , Proteínas Morfogenéticas Ósseas/fisiologia , Epitélio/fisiologia , Masculino , Alvéolos Pulmonares/patologia , Alvéolos Pulmonares/fisiologia , Enfisema Pulmonar/patologia , Enfisema Pulmonar/fisiopatologia , Ratos , Ratos Sprague-Dawley , Regeneração/fisiologia , Transdução de Sinais/fisiologia , Fator de Crescimento Transformador beta/fisiologia , Tretinoína/farmacologia , Vimentina/genéticaRESUMO
INTRODUCTION: Pulmonary emphysema characterized by alveolar wall destruction is resultant of persistent chronic inflammation. All-trans retinoic acid (ATRA) has been reported to reverse elastase-induced emphysema in rats. However, the underlying molecular mechanisms are so far unknown. OBJECTIVE: To investigate the therapeutic potential effect of ATRA via the amelioration of the ERK/JAK-STAT pathways in the lungs of emphysematous rats. METHODS: In silico analysis was done to find the binding efficiency of ATRA with receptor and ligands of ERK & JAK-STAT pathway. Emphysema was induced by porcine pancreatic elastase in Sprague-Dawley rats and ATRA was supplemented as therapy. Lungs were harvested for histopathological, genomics and proteomics analysis. RESULTS AND DISCUSSION: In silico docking, analysis confirms that ATRA interferes with the normal binding of ligands (TNF-α, IL6ST) and receptors (TNFR1, IL6) of ERK/JAK-STAT pathways respectively. ATRA restored the histology, proteases/antiproteases balance, levels of inflammatory markers, antioxidants, expression of candidate genes of ERK and JAK-STAT pathways in the therapy group. CONCLUSION: ATRA ameliorates ERK/JAK-STAT pathway in emphysema condition, resulting in alveolar epithelium regeneration. Hence, ATRA may prove to be a potential drug in the treatment of emphysema.
Assuntos
Anti-Inflamatórios/farmacologia , Alvéolos Pulmonares/efeitos dos fármacos , Enfisema Pulmonar/tratamento farmacológico , Regeneração/efeitos dos fármacos , Tretinoína/farmacologia , Animais , Janus Quinases/fisiologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Elastase Pancreática/metabolismo , Elastase Pancreática/farmacologia , Alvéolos Pulmonares/fisiologia , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição STAT/fisiologia , Tretinoína/uso terapêutico , Fator de Necrose Tumoral alfa/análiseAssuntos
Oxigenoterapia Hiperbárica/enfermagem , Humanos , Oxigenoterapia Hiperbárica/efeitos adversos , Oxigenoterapia Hiperbárica/instrumentação , Masculino , Pessoa de Meia-Idade , Fenômenos Físicos , Alvéolos Pulmonares/fisiologia , Medição de Risco , Terminologia como Assunto , Resultado do TratamentoRESUMO
Resistance respiratory muscle training (RRMT) increases respiratory muscle strength and can increase swimming endurance time by as much as 85%. The purpose of this study was to examine potential mechanisms by which RRMT improves exercise endurance. Eight healthy adult male scuba divers underwent experiments in a hyperbaric chamber at sea level (1 atmosphere absolute (ATA)), 2.7 ATA and 4.6 ATA, both dry and fully submersed. Subjects rested, exercised, and rested while mimicking their own exercise breathing (ISEV). Airway resistance (R(aw)), exhaled nitric oxide output (VË(NO)), and respiratory duty cycle (T(I)/T(Tot)) were determined before and after four weeks of RRMT. RRMT decreased T(I)/T(Tot) (-10% at rest at 1 ATA), VË(O2) (-17% at 2.7 ATA during submersed exercise), VË(E) (-6% at 2.7 ATA during submersed exercise), and R(aw) (-34% inspiratory at 4.6 ATA submersed, -38% expiratory at 2.7 ATA dry), independent of changes in VË(NO). Most importantly, respiratory muscle efficiency increased (+83% at 2.7 ATA submersed).
Assuntos
Exercícios Respiratórios/métodos , Mergulho/fisiologia , Esforço Físico/fisiologia , Mecânica Respiratória/fisiologia , Músculos Respiratórios/fisiologia , Adulto , Resistência das Vias Respiratórias/fisiologia , Humanos , Masculino , Óxido Nítrico/metabolismo , Consumo de Oxigênio/fisiologia , Resistência Física/fisiologia , Pressão , Alvéolos Pulmonares/fisiologia , Descanso/fisiologia , Resultado do TratamentoRESUMO
ß-Carotene 15,15'-monooxygenase (CMO1, BCMO1) converts ß-carotene to retinaldehyde (retinal) and is a key enzyme in vitamin A metabolism. CMO1 activity is robust in the intestine and liver, where cmo1 gene transcription may be subject to negative feedback by accumulation of its metabolic products. Evidence from CMO1 null animals also indicates that non-gastrointestinal CMO1 may be required for tissue-specific conversion of ß-carotene into vitamin A. The aim of this study was to investigate the effects of the enzymatic substrate, ß-carotene, on regulation of CMO1 in a cell model of human alveolar pneumocytes. We demonstrate that CMO1 is expressed in human alveolar epithelial (A549) cells and converts ß-carotene into retinal and biologically active retinoic acids (RA). Exposure to ß-carotene suppresses CMO1 expression at both mRNA and protein levels. ß-Carotene, but not all-trans RA, decreases CMO1 promoter activity in a time- and dosage-dependent manner. This ß-carotene-mediated inhibition of CMO1 expression results from decreased binding of peroxisome proliferator-activated receptor γ (PPARγ) and retinoid X receptor α (RXRα) in the CMO1 promoter. ß-Carotene treatment also antagonizes PPARγ activity in HEK293 cells that stably express CMO1 wild-type, but not in cells that express the CMO1 mutant or vector alone. These findings have implications for local vitamin A synthesis in the lung, especially during systemic vitamin A insufficiency and may also help to explain, in part, the mechanism underlying the increased lung cancer risk upon ß-carotene supplementation in smokers.
Assuntos
Regulação Enzimológica da Expressão Gênica , Alvéolos Pulmonares/enzimologia , Mucosa Respiratória/metabolismo , beta Caroteno/fisiologia , beta-Caroteno 15,15'-Mono-Oxigenase/genética , Linhagem Celular Tumoral , Regulação para Baixo/genética , Células HEK293 , Humanos , PPAR gama/antagonistas & inibidores , PPAR gama/genética , PPAR gama/metabolismo , Ligação Proteica/genética , Alvéolos Pulmonares/patologia , Alvéolos Pulmonares/fisiologia , Mucosa Respiratória/enzimologia , Mucosa Respiratória/patologia , Regulação para Cima/genética , beta Caroteno/metabolismo , beta-Caroteno 15,15'-Mono-Oxigenase/antagonistas & inibidores , beta-Caroteno 15,15'-Mono-Oxigenase/biossínteseRESUMO
BACKGROUND: Pulmonary alveolar proteinosis (PAP) is a syndrome with multiple etiologies and is often deadly in lysinuric protein intolerance (LPI). At present, PAP is treated by whole lung lavage or with granulocyte/monocyte colony stimulating factor (GM-CSF); however, the effectiveness of GM-CSF in treating LPI associated PAP is uncertain. We hypothesized that GM-CSF and surfactant protein D (SP-D) would enhance the clearance of proteins and dying cells that are typically present in the airways of PAP lungs. METHODS: Cells and cell-free supernatant of therapeutic bronchoalveolar lavage fluid (BALF) of a two-year-old patient with LPI were isolated on multiple occasions. Diagnostic BALF samples from an age-matched patient with bronchitis or adult PAP patients were used as controls. SP-D and total protein content of the supernatants were determined by BCA assays and Western blots, respectively. Cholesterol content was determined by a calorimetic assay or Oil Red O staining of cytospin preparations. The cells and surfactant lipids were also analyzed by transmission electron microscopy. Uptake of Alexa-647 conjugated BSA and DiI-labelled apoptotic Jurkat T-cells by BAL cells were studied separately in the presence or absence of SP-D (1 microg/ml) and/or GM-CSF (10 ng/ml), ex vivo. Specimens were analyzed by light and fluorescence microscopy. RESULTS: Here we show that large amounts of cholesterol, and large numbers of cholesterol crystals, dying cells, and lipid-laden foamy alveolar macrophages were present in the airways of the LPI patient. Although SP-D is present, its bioavailability is low in the airways. SP-D was partially degraded and entrapped in the unusual surfactant lipid tubules with circular lattice, in vivo. We also show that supplementing SP-D and GM-CSF increases the uptake of protein and dying cells by healthy LPI alveolar macrophages, ex vivo. Serendipitously, we found that these cells spontaneously generated granulomas, ex vivo, and GM-CSF treatment drastically increased the number of granulomas whereas SP-D treatment counteracted the adverse effect of GM-CSF. CONCLUSIONS: We propose that increased GM-CSF and decreased bioavailability of SP-D may promote granuloma formation in LPI, and GM-CSF may not be suitable for treating PAP in LPI. To improve the lung condition of LPI patients with PAP, it would be useful to explore alternative therapies for increasing dead cell clearance while decreasing cholesterol content in the airways.
Assuntos
Erros Inatos do Metabolismo dos Aminoácidos/terapia , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Granuloma/metabolismo , Lisina/metabolismo , Macrófagos Alveolares/fisiologia , Proteína D Associada a Surfactante Pulmonar , Adulto , Erros Inatos do Metabolismo dos Aminoácidos/imunologia , Erros Inatos do Metabolismo dos Aminoácidos/metabolismo , Erros Inatos do Metabolismo dos Aminoácidos/fisiopatologia , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Pré-Escolar , Colesterol/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/uso terapêutico , Humanos , Pulmão/citologia , Pulmão/metabolismo , Proteinose Alveolar Pulmonar/imunologia , Proteinose Alveolar Pulmonar/metabolismo , Proteinose Alveolar Pulmonar/fisiopatologia , Proteinose Alveolar Pulmonar/terapia , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/fisiologia , Proteína D Associada a Surfactante Pulmonar/metabolismo , Proteína D Associada a Surfactante Pulmonar/uso terapêutico , Resultado do TratamentoRESUMO
BACKGROUND: The optimal oxygen concentration for the resuscitation of premature infants remains controversial. OBJECTIVES: We studied the effects of 21 versus 100% oxygen at initial resuscitation and also the effects of 24-hour exposure to 100% oxygen on arterial blood gases, oxidant lung injury, activities of lung antioxidant enzymes (AOEs) and isolated pulmonary artery (PA) contractility in preterm newborn lambs. METHODS: Preterm lambs at 128 days' gestation (term = 145 days) were delivered and ventilated with 21 (RAR; n = 5) or 100% oxygen (OXR; n = 5) for the first 30 min of life. Subsequently, FiO2 was adjusted to maintain an arterial PO2 (PaO2) between 45 and 70 mm Hg for 24 h. A third group of lambs was mechanically ventilated with 100% oxygen for 24 h (OX24; n = 5). RESULTS: Oxidized glutathione levels in whole blood correlated highly with PaO2. Reduced to oxidized glutathione ratio was significantly different between the groups, the ratio increasing with decreasing oxygen exposure. The OX24 group had significantly higher activities of lipid hydroperoxide and myeloperoxidase and significantly lower activities of superoxide dismutase, catalase and glutathione peroxidase in the lung at 24 h. Activities of AOEs correlated inversely with alveolar PO2. PA contractility to norepinephrine and KCl was greater with increasing oxygen exposure. Pretreatment with superoxide dismutase and catalase significantly reduced PA contractility in the OXR and OX24 groups, but not in the RAR group. CONCLUSIONS: We conclude that ventilated premature lambs are unable to appropriately increase AOE activity in response to hyperoxia and that increasing exposure to oxygen aggravates systemic oxidant stress, oxidant lung injury and pulmonary arterial contractility in these lambs.
Assuntos
Pulmão/efeitos dos fármacos , Oxirredutases/metabolismo , Oxigênio/administração & dosagem , Artéria Pulmonar/efeitos dos fármacos , Vasoconstrição/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Catalase/metabolismo , Catalase/farmacologia , Modelos Animais de Doenças , Regulação para Baixo , Interações Medicamentosas , Idade Gestacional , Dissulfeto de Glutationa/sangue , Técnicas In Vitro , Peróxidos Lipídicos/metabolismo , Pulmão/enzimologia , Norepinefrina/farmacologia , Oxigênio/sangue , Cloreto de Potássio/farmacologia , Alvéolos Pulmonares/fisiologia , Artéria Pulmonar/enzimologia , Respiração Artificial , Ressuscitação , Ovinos , Superóxido Dismutase/metabolismo , Superóxido Dismutase/farmacologia , Vasoconstrição/fisiologiaRESUMO
We showed previously that removing 55-58% of the lung by right pneumonectomy (R-PNX) in adult dogs triggers compensatory growth of the remaining lung, but removing 42-45% of the lung by left PNX (L-PNX) does not. We also showed that, following R-PNX, supplemental all-trans retinoic acid (RA) selectively enhances alveolar capillary endothelial cell volume (Yan X, Bellotto DJ, Foster DJ, Johnson RL, Jr., Hagler HH, Estrera AS, and Hsia CC. J Appl Physiol 96: 1080-1089, 2004). We hypothesized that RA supplementation might enhance compensation following L-PNX and tested this hypothesis by administering RA (2 mg.kg(-1).day(-1), 4 days/wk) or placebo orally to litter-matched adult foxhounds for 4 mo following L-PNX. Resting lung function was measured under anesthesia. Air and tissue volumes of the remaining lung were assessed by high-resolution computed tomography scan and by detailed postmortem morphometric analysis of the fixed lung. There was no significant difference in resting lung function, lung volume, alveolar structure, or septal ultrastructure between RA and placebo treatment groups. We conclude that RA supplementation does not induce post-PNX compensatory lung growth in the absence of existing cellular growth activities initiated by other primary signals.
Assuntos
Antineoplásicos/farmacologia , Pneumonectomia , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/crescimento & desenvolvimento , Tretinoína/farmacologia , Adaptação Fisiológica/efeitos dos fármacos , Animais , Cães , Medidas de Volume Pulmonar , Masculino , Microcirculação/efeitos dos fármacos , Alvéolos Pulmonares/fisiologia , Circulação Pulmonar/efeitos dos fármacos , Estresse Mecânico , Tomografia Computadorizada por Raios XRESUMO
Undersulfation of the basement membrane matrix of alveolar type II (AT2) cells compared with that of neighboring type I cells is believed to account for some of the known morphological and functional differences between these pneumocytes. Heparin, a model for sulfated components of basement membrane matrices, is known to inhibit fibroblast growth factor (FGF)-2-stimulated DNA synthesis as well as gene expression of FGF-2 and its receptor in AT2 cells. To determine whether these end points result from specific effects of heparin on FGF-related signaling pathways, isolated rat AT2 cells were treated with 100 ng/ml FGF-1 or FGF-2 in the presence of up to 500 microg/ml heparin. In addition, experiments were done on cells grown in the presence of 20 mM sodium chlorate (sulfation inhibitor). High-dose heparin reduced FGF-1- or FGF-2-stimulated phosphorylation of mitogen-activated protein kinase kinases (MEK1/2), p44/42 mitogen-activated protein kinases (MAPK/ERK1/2), stress-activated protein kinase/c-Jun NH(2)-terminal kinase, Akt/protein kinase B, and p90(RSK). FGF-2-stimulated signaling was more sensitive to heparin's effects than was signaling stimulated by FGF-1. Heparin had an additive effect on the reduced [(3)H]thymidine incorporation in FGF-2-treated AT2 cells caused by inhibition of the MEK/ERK pathway by the MEK inhibitor PD-98059. The data suggest that heparin's known capacity to alter DNA synthesis and, possibly, other biological end points is realized via cross talk between multiple signaling pathways.
Assuntos
Fator 1 de Crescimento de Fibroblastos/farmacologia , Fator 2 de Crescimento de Fibroblastos/farmacologia , Heparina/farmacologia , Proteínas Serina-Treonina Quinases , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/fisiologia , Transdução de Sinais/efeitos dos fármacos , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Heparina/administração & dosagem , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/metabolismo , Ratos , Ratos Endogâmicos , Sulfatos/metabolismo , Timidina/metabolismo , Fatores de TempoRESUMO
Airway hyperresponsiveness (AHR) is influenced by structural components of the bronchial wall, including the smooth muscle and connective tissue elements and the neuromuscular function. AHR is also influenced by parenchymally derived tethering forces on the bronchial wall, which maintain airway caliber by producing outward radial traction. Our previous work has shown that vitamin A-deficient (VAD) rats exhibit cholinergic hyperresponsiveness and a decrease in the expression and function of the muscarinic-2 receptors (M2R). We hypothesized that if decreases in radial traction from airway or parenchymal structures contributed to the VAD-related increase in AHR, then the radial traction would normalize more slowly than VAD-related alterations in neurotransmitter signaling. Rats remained vitamin A sufficient (VAS) or were rendered VAD and then maintained on the VAD diet in the presence or absence of supplementation with all-trans retinoic acid (RA). VAD was associated with an approximately twofold increase in respiratory resistance and elastance compared with VAS rats. Exposure to RA for 12 days but not 4 days restored resistance and elastance to control (VAS) levels. In VAD rats, AHR was accompanied by decreases in bronchial M2R gene expression and function, which were restored after 12 days of RA supplementation. Subepithelial bronchial elastic fibers were decreased by approximately 50% in VAD rats and were significantly restored by RA. The increase in AHR that is associated with VAD is accompanied by decreases in M2R expression and function that can be restored by RA and a reduction in airway elastic fibers that can be partially restored by RA.
Assuntos
Antineoplásicos/farmacologia , Hiper-Reatividade Brônquica/tratamento farmacológico , Hiper-Reatividade Brônquica/etiologia , Tretinoína/farmacologia , Deficiência de Vitamina A/complicações , Deficiência de Vitamina A/fisiopatologia , Animais , Broncoconstritores/farmacologia , Elasticidade , Feminino , Expressão Gênica , Cloreto de Metacolina/farmacologia , Agonistas Muscarínicos/farmacologia , Pilocarpina/farmacologia , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/fisiologia , Ratos , Ratos Endogâmicos Lew , Receptor Muscarínico M2/genética , Receptor Muscarínico M2/metabolismo , Organismos Livres de Patógenos EspecíficosRESUMO
The middle-sized bronchus constricts during mid-inspiration through early-expiration. The purpose of this study was to elucidate the physiological role of this respiratory-related bronchial rhythmic constriction (RRBRC). The following parameters were measured in 12 decerebrated and paralyzed dogs: pressure from a balloon-tipped catheter in the fifth-generation bronchus (to reveal RRBRC), efferent neurogram from C(5) phrenic, and ventilatory flow and volume. We found a small but significant reduction of peak expiratory flow of mechanical ventilation during RRBRC. During bilateral vagal cold block, RRBRC was simulated by intermittent electric stimulation of vagal fibers distal to the cold block. This stimulus evoked a decrease in peak expiratory flow and in Pa(CO2) (approximately 1.5 mmHg). After vagal warming, mechanical ventilation was terminated, and blood gases were maintained normal by extracorporeal oxygenation. During each RRBRC ventilatory volume decreased by approximately 3 ml. The changes in gas volume and RRBRC disappeared after bilateral vagotomy. These findings support the concept that the physiological role of RRBRC is to facilitate alveolar gas exchange by reducing expiratory flow, anatomical dead space, or both.
Assuntos
Brônquios/fisiologia , Broncoconstrição/fisiologia , Alvéolos Pulmonares/fisiologia , Ventilação Pulmonar/fisiologia , Respiração , Potenciais de Ação , Animais , Gasometria , Pressão Sanguínea , Dióxido de Carbono/metabolismo , Temperatura Baixa , Cães , Relação Dose-Resposta à Radiação , Estimulação Elétrica , Fármacos Neuromusculares não Despolarizantes , Oxigênio/metabolismo , Pancurônio , Nervo Frênico/fisiologia , Respiração Artificial/métodos , Paralisia Respiratória/induzido quimicamente , Paralisia Respiratória/fisiopatologia , Volume de Ventilação Pulmonar/fisiologia , Vagotomia/métodos , Nervo Vago/fisiopatologiaRESUMO
Oxidants play an important role in the development of acute and chronic lung injuries. Alveolar surfactant is the first target of air-borne oxidants. Surfactant contains, besides dipalmitoyl phosphatidylcholine, cholesterol and polyunsaturated phospholipids that play an important functional role. Therefore, vitamin E could be important for protecting surfactant lipids against oxidation and subsequent lung injury. Alveolar type II cells play a central role in synthesis and secretion of surfactant lipids and also supplement the surfactant with vitamin E during intracellular assembly. High density lipoprotein (HDL) is the primary source of vitamin E for type II cells. The uptake of vitamin E by specific lipid transfer is mediated by at least three HDL-specific receptors (scavenger receptor BI, membrane dipeptidase, and HDL-binding protein-2). In addition, cubilin and megalin mediate in a cooperative manner HDL-holoparticle uptake by alveolar type II cells. A temporary vitamin E deficiency induces a reversible change of the expression of pro- and antiinflammatory markers and of markers defining apoptosis, and reduces surfactant lipid synthesis in alveolar type II cells. These metabolic changes of type II cells may prime the lung to develop clinically manifest injury in response to an additional insult, e.g., hyperoxia.
Assuntos
Antioxidantes/metabolismo , Proteínas de Membrana , Alvéolos Pulmonares/fisiologia , Receptores Imunológicos/fisiologia , Receptores de Lipoproteínas , Deficiência de Vitamina E/metabolismo , Vitamina E/fisiologia , Animais , Humanos , Alvéolos Pulmonares/metabolismo , Receptores Imunológicos/metabolismo , Receptores Depuradores , Receptores Depuradores Classe B , Vitamina E/metabolismoRESUMO
By EPR spectroscopy, we investigated free radical production by cultured human alveolar cells subjected to anoxia/re-oxygenation (A/R), and tested the effects of ceftazidime, an antibiotic previously demonstrated to possess antioxidant properties. Two A/R models were performed on type II pneumocytes (A549 cell line), either on cells attached to culture dishes (monolayer A/R model; 3.5 h of anoxia, 30 min of re-oxygenation) or after cell detachment (suspension A/R model; 1 h of anoxia, 10 min of re-oxygenation). Ceftazidime and selective inhibitors (SOD, Tiron, L-NMMA) were added before anoxia. Free radical production was assessed by the EPR spin trapping technique. Oxygen consumption was monitored, in parallel with EPR studies, in the suspension A/R model. The production of free radical species was demonstrated by the generation of PBN-radical adducts: (a(N) = 15.2 G) in the monolayer A/R model and a six-line EPR spectrum (a(N) = 15.7 G and a(H) = 2.7 G) in the suspension A/R model. A kinetic study performed by oximetry, in parallel with EPR spectroscopy, demonstrated marked alterations of the cell respiratory function and that the free radical production started during anoxia and increased during re-oxygenation. In the suspension A/R model, the amplitude of EPR spectra were decreased upon the addition of 200 U/ml SOD (37% inhibition), 0.1 mM Tiron (67% inhibition) and 1 mM L-NMMA (43% inhibition). Addition of 1 mM ceftazidime decreased the amplitude of EPR spectra (37% inhibition) in both A/R models. Complementary in vitro EPR studies demonstrated that CAZ scavenged the hydroxyl radical (produced by the Fenton reaction). The protective effect of ceftazidime in the cell model could thus be linked to its ability to scavenge superoxide anions, nitrogen-derived species and hydroxyl radicals.
Assuntos
Ceftazidima/farmacologia , Hipóxia Celular/fisiologia , Consumo de Oxigênio , Alvéolos Pulmonares/fisiologia , Adenocarcinoma , Antibacterianos/farmacologia , Hipóxia Celular/efeitos dos fármacos , Espectroscopia de Ressonância de Spin Eletrônica/métodos , Radicais Livres/metabolismo , Humanos , Neoplasias Pulmonares , Consumo de Oxigênio/efeitos dos fármacos , Alvéolos Pulmonares/efeitos dos fármacos , Células Tumorais Cultivadas , ômega-N-Metilarginina/farmacologiaRESUMO
In order to respond to the flood of new active ingredients currently being generated by combinatorial chemistry or molecular biological synthesis, selection procedures able to filter out rapidly and economically those drug candidates with the highest development potential are required. This necessitates the measurement of fundamental biopharmaceutical parameters very early in the drug development process. Any pharmaceutically active agent must be able to overcome the body's natural protective mechanisms. A broad variety of biological barriers can be simulated in the laboratory by cell monolayer models. Apart from ethical aspects, the advantage of these in vitro test systems is that permeability studies can be performed at high throughput rates under controlled and reproducible conditions. The validity of such a model is ultimately reflected in its ability to accurately predict the behaviour of an active ingredient at the corresponding in vivo barrier.
Assuntos
Barreira Hematoencefálica , Fenômenos Fisiológicos do Sistema Digestório , Avaliação Pré-Clínica de Medicamentos/métodos , Preparações Farmacêuticas/metabolismo , Alvéolos Pulmonares/fisiologia , Mucosa Respiratória/fisiologia , Alternativas aos Testes com Animais , Animais , Técnicas de Cultura de Células/métodos , Linhagem Celular , Permeabilidade da Membrana Celular , Humanos , Modelos Biológicos , Células Tumorais CultivadasRESUMO
Chest physiotherapy in connection with abdominal surgery includes different deep-breathing exercises to prevent post-operative pulmonary complications. The therapy is effective in preventing pulmonary complications, especially in high-risk patients such as obese persons. The mechanisms behind the effect is unclear, but part of the effect may be explained by the changes in breathing patterns. The aim of this study was therefore to describe and to analyse the breathing patterns in obese and non-obese subjects during three different breathing techniques frequently used in the treatment of post-operative patients. Twenty-one severely obese [body mass index (BMI) > 40] and 21 non-obese (BMI 19-25) subjects were studied. All persons denied having any lung disease and were non-smokers. The breathing techniques investigated were: deep breaths without any resistance (DB), positive expiratory pressure (PEP) with an airway resistance of approximately +15 cmH2O (1.5 kPa) during expiration, inspiratory resistance positive expiratory pressure (IR-PEP) with a pressure of approximately -10 cmH2O (-1.0 kPa) during inspiration. Expiratory resistance as for PEP. Volume against time was monitored while the subjects were sitting in a body plethysmograph. Variables for volume and flow during the breathing cycle were determined. Tidal volume and alveolar ventilation were highest during DB, and peak inspiratory volume was significantly higher than during PEP and IR-PEP in the group of obese subjects. The breathing cycles were prolonged in all techniques but were most prolonged in PEP and IR-PEP. The functional residual capacity (FRC) was significantly lower during DB than during PEP and IR-PEP in the group of obese subjects. FRC as determined within 2 min of finishing each breathing technique was identical to before the breathing manoeuvres.
Assuntos
Exercícios Respiratórios , Obesidade/fisiopatologia , Obesidade/terapia , Adulto , Resistência das Vias Respiratórias , Feminino , Fluxo Expiratório Forçado , Capacidade Residual Funcional , Humanos , Masculino , Pletismografia , Pressão , Alvéolos Pulmonares/fisiologia , Mecânica Respiratória/fisiologia , Capacidade Pulmonar TotalRESUMO
Utilizing the sulfur-containing gases of garlic as probes, we investigated the gut versus mouth origin of odoriferous breath gases. Five individuals ingested 6 g of garlic, and sulfur gases in mouth, alveolar air, and urine samples were measured. The mouth normally contained low concentrations of hydrogen sulfide, methanethiol, and dimethyl sulfide. Immediately after garlic ingestion, transient high concentrations of methanethiol and allyl mercaptan and lesser concentrations of allyl methyl sulfide (AMS), allyl methyl disulfide, and allyl disulfide were observed. With the exception of AMS, all gases were present in far greater concentrations in mouth than alveolar air, indicating an oral origin. Only AMS was of gut origin as evidenced by similar partial pressures in mouth, alveolar air, and urine. After 3 h, AMS was the predominant breath sulfur gas. The unique derivation of AMS from the gut is attributable to the lack of gut and liver metabolism of this gas versus the rapid metabolism of the other gases. Breath odor after garlic ingestion initially originates from the mouth and subsequently from the gut.
Assuntos
Ingestão de Alimentos , Alho , Halitose/etiologia , Intestinos/fisiologia , Boca/fisiologia , Plantas Medicinais , Adulto , Ingestão de Alimentos/fisiologia , Feminino , Gases , Humanos , Mucosa Intestinal/metabolismo , Masculino , Pessoa de Meia-Idade , Boca/metabolismo , Concentração Osmolar , Alvéolos Pulmonares/fisiologia , Compostos de Enxofre/metabolismo , Compostos de Enxofre/urinaRESUMO
Hypoxic pulmonary vasoconstriction underlies the development of high-altitude pulmonary edema. Anecdotal observations suggest a beneficial effect of garlic in preventing high-altitude symptoms. To determine whether garlic influences pulmonary vasoconstriction, we assessed the effect of garlic on pulmonary pressures in rats subjected to alveolar hypoxia and on vasoconstriction in isolated pulmonary arterial rings. Garlic gavage (100 mg/kg body wt) for 5 days resulted in complete inhibition of acute hypoxic pulmonary vasoconstriction compared with the control group. No difference in mean arterial pressure or heart rate response to hypoxia was seen between the groups. Garlic solution resulted in a significant dose-dependent vasorelaxation in both endothelium-intact and mechanically endothelium-disrupted pulmonary arterial rings. The administration of NG-nitro-L-arginine methyl ester (a nitric oxide synthase inhibitor) inhibited the vasodilatory effect of garlic by 80%. These studies document that garlic blocks hypoxic pulmonary hypertension in vivo and demonstrate a combination of endothelium-dependent and -independent mechanisms for the effect in pulmonary arterial rings.
Assuntos
Alho/uso terapêutico , Hipertensão Pulmonar/prevenção & controle , Hipóxia , Músculo Liso Vascular/fisiologia , Fitoterapia , Extratos Vegetais/farmacologia , Plantas Medicinais , Artéria Pulmonar/fisiologia , Acetilcolina/farmacologia , Animais , Pressão Sanguínea , Endotélio Vascular/fisiologia , Hipertensão Pulmonar/fisiopatologia , Técnicas In Vitro , Masculino , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/fisiopatologia , NG-Nitroarginina Metil Éster/farmacologia , Fenilefrina/farmacologia , Alvéolos Pulmonares/fisiologia , Alvéolos Pulmonares/fisiopatologia , Artéria Pulmonar/efeitos dos fármacos , Artéria Pulmonar/fisiopatologia , Ratos , Ratos Sprague-Dawley , VasoconstriçãoRESUMO
We evaluated four ventilator patterns after the administration of 80 mg/kg bovine lipid extract surfactant (LES) to anesthetized, paralyzed, saline-lavaged New Zealand white rabbits. Two ventilator types were compared: high frequency oscillatory ventilation (HFO) versus conventional mechanical ventilation (CMV), each at high (HI) and low (LO) end-expiratory lung volumes (EELV); n = 6, each group; treatment duration = 4 h. Target PaO2 ranges were > 350 mm Hg for groups with high EELV (i.e., HFO-HI and CMV-HI) and 70 to 100 mm Hg for those with low EELV (i.e., HFO-LO and CMV-LO). Ventilator pressures were limited to < or = 39/9 cm H2O in the CMV-HI group. Five of six CMV-HI-treated animals did not maintain target PaO2 levels. Both ventilator type and strategy influenced outcome significantly. Animals managed with HFO had higher mean arterial pressures (p = 0.004), lower mean airway pressures (Paw) (p < 0.00008) and HCO3- requirements (p < 0.02), larger inflation (p = 0.003) and deflation (p < 0.00001) respiratory system volumes at 10 cm inflation pressure, and higher lung lamellar body (p = 0.0006) and lavage fluid (p = 0.003) phospholipid quantities than did CMV-treated animals. The deflation P-V curve (p = 0.0004), lamellar body (p < 0.00001) and lavage fluid (p = 0.0002) phospholipid levels were superior after the high EELV strategy. We conclude that ventilator pattern strongly influences exogenous surfactant efficacy. Benefits arise from keeping EELV high enough to prevent atelectasis and using small (approximately 2 ml/kg) tidal volumes to prevent overdistension.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Alvéolos Pulmonares/fisiologia , Surfactantes Pulmonares/uso terapêutico , Respiração Artificial/métodos , Análise de Variância , Animais , Líquido da Lavagem Broncoalveolar/química , Avaliação Pré-Clínica de Medicamentos , Pulmão/química , Fosfolipídeos/análise , Troca Gasosa Pulmonar , Surfactantes Pulmonares/deficiência , Coelhos , Distribuição Aleatória , Respiração Artificial/instrumentação , Respiração Artificial/estatística & dados numéricos , Mecânica Respiratória , Fatores de TempoRESUMO
Potential toxic effects of prolonged NO2 exposure below the current threshold limit value (TLV) were examined in 14 healthy, nonsmoking adults. The subjects were exposed to 2.3 ppm NO2 and to clean air for 5 h with a 1-wk interval between exposures. Physiologic and biochemical measurements were obtained during the exposures and until 24 h after. A 14% decrease in serum glutathione peroxidase activity (GSH-Px) was observed 24 h after the start of the NO2 exposure, while indications of a 22% decrease in alveolar permeability were found 11 h after the start. There were no indications of mucous membrane irritation or of decreased lung function during or after NO2 exposures. The results support the assumption that a delayed response is a feature of the human reaction to NO2 even below the current TLV of 3 ppm, and they stress the importance of an extended period of observation in future NO2 exposure studies.
Assuntos
Permeabilidade da Membrana Celular/efeitos dos fármacos , Dióxido de Nitrogênio/toxicidade , Alvéolos Pulmonares/efeitos dos fármacos , Adulto , Análise de Variância , Câmaras de Exposição Atmosférica , Permeabilidade da Membrana Celular/fisiologia , Método Duplo-Cego , Feminino , Glutationa/sangue , Glutationa Peroxidase/sangue , Glutationa Peroxidase/efeitos dos fármacos , Humanos , Masculino , Alvéolos Pulmonares/fisiologia , Testes de Função Respiratória/estatística & dados numéricos , Selênio/sangue , Fatores de TempoRESUMO
To reveal the regulatory mechanism of the mineralocorticoid aldosterone on basolateral K+ channels, the aldosterone-sensitive lung epithelium of Xenopus laevis was investigated in Ussing chambers under voltage-clamp conditions. Transepithelial measurements were supplemented by current fluctuation analysis of short-circuit current noise in nonstimulated and aldosterone-stimulated lung tissues. The addition of 10(-6) M aldosterone stimulated short-circuit current from 11.3 +/- 2.0 to 27.8 +/- 4.8 microA/cm2 (n = 11) within 4-5 h. In the presence of an alveolar-to-pleural K+ gradient, transepithelial K+ currents were induced by permeabilizing the apical membrane with the pore-forming antibiotic amphotericin B. When the local anesthetic lidocaine (25-1,000 microM) was added to the pleural solution, macroscopic K+ current was dose dependently depressed. Lidocaine induced a Lorentzian component in the power density spectra, and the corner frequency increased linearly with blocker concentration. Aldosterone treatment did not affect mean single K+ channel current, which was 1.5 +/- 0.12 pA corresponding to a 15-pS channel conductance, whereas the number of basolateral K+ channels doubled. We conclude that the basolateral K+ channels in alveolar epithelia are a target site of aldosterone action.