RESUMO
The aim of this study was to analyze direct costs of COPD therapy in relation with clinical course and stage of the disease. Sixty patients with moderate to severe COPD were included into the study. The average cost was taken from institutional data file and was also assessed from a social perspective. Results were presented as average costs per patient per year. Forty two percent of patients was classified as GOLD D category, while categories A, B, and C accounted for 8 %, 27 %, and 23 %, respectively. Approximately 65 % of patients had 2-3 degrees of dyspnea according to the Modified Medical Research Council Dyspnea Scale. About 60 % of patients underwent two or three exacerbations per year and those patients had one or two co-morbidities diagnosed. Treatment costs almost doubled with disease progression, mainly due to exacerbations. In patients in Group C and Group D with exacerbations the direct costs were several times higher than in group A or B and the difference increased with progression of the disease. In Groups A and B, the costs of treatment of stable disease or with exacerbation were comparable. We conclude that costs of treatment of COPD patients were highest in advanced disease and were strongly related to COPD exacerbations.
Assuntos
Custos de Cuidados de Saúde , Doença Pulmonar Obstrutiva Crônica/economia , Idoso , Volume Expiratório Forçado , Humanos , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/fisiopatologiaRESUMO
UNLABELLED: Observational studies have suggested that statins may have beneficial effects on outcomes in chronic obstructive pulmonary disease (COPD) patients. These effects may be mediated through an anti-inflammatory effect of statins. The purpose of this pilot-study was to determine whether statins have an anti-inflammatory effect on the lungs of COPD patients. We conducted randomized, controlled, parallel group pilot-study to compare the effects of atorvastatin (n=12) or placebo (n=6) on lung inflammation in patients with mild to moderate COPD. The primary endpoint was change in CD45+ cells expression measured by immunohistochemistry and changes in expression of genes measured using microarrays in lung biopsy (TBB) samples before and after 12 weeks of treatment with atorvastatin 40 mg/day. All subjects had spirometry, lung volumes, diffusing capacity of the lungs for carbon monoxide (DLCO), St George's Respiratory Questionnaire (SGRQ), 6 minute walk distance (6 MWD), serum lipids, hs-CRP, induced sputum (IS), bronchoscopy and TBB carried out at baseline and after treatment. TBB specimens were processed for histology, immunohistochemistry and genome-wide association studies (GWAS) profiling. Seventeen subjects completed the study. There was a significant improvement in SGRQ with mean SGRQ decreased by 12 points after treatment with atorvastatin (P=0.012). Atorvastatin treatment produced a significant 34% reduction in sputum neutrophil count, and a 57% reduction in CD45+ cells in lung biopsies (expressed as integrated optical density -IOD; median IOD 62.51% before, 27.01% after atorvastatin treatment, P=0.008). In patients' lung tissue atorvastatin treatment produced downregulation of key genes involved in inflammatory processes, immune response, and leukocyte activation. These data demonstrate the pulmonary anti-inflammatory effects of atorvastatin in COPD patients with the potential for beneficial clinical effects. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01748279.
Assuntos
Anti-Infecciosos/uso terapêutico , Atorvastatina/uso terapêutico , Pulmão/efeitos dos fármacos , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Idoso , Biópsia , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Marcadores Genéticos , Estudo de Associação Genômica Ampla , Humanos , Imuno-Histoquímica , Mediadores da Inflamação/imunologia , Pulmão/imunologia , Pulmão/fisiopatologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Polônia , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/imunologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Recuperação de Função Fisiológica , Testes de Função Respiratória , Índice de Gravidade de Doença , Método Simples-Cego , Fatores de Tempo , Resultado do TratamentoRESUMO
Immune cells expressing the activation markers HLA-DR and regulatory T cells (Tregs) may be involved in the regulation of chronic inflammation in chronic obstructive pulmonary disease (COPD). In this study we analyzed native and activated cell profiles in sputum of 22 stable COPD patients receiving formoterol (F) or formoterol + tiotropium (F + T) for 3 months. Cells were isolated from induced sputum and were examined on Coulter flow cytometer using fluorescent antibodies specific for CD3, CD4, CD8, CD14, CD19, CD25, CD127, and HLA-DR antigens. Cell profiles and cell activation were assessed by analysis of HLA-DR, CD25, and CD127 co-expression in double-stained samples. Tregs were defined as CD4âºCD25(high) CD127(low) cells. We found that the combined therapy significantly decreased the CD8⺠cell number (p < 0.01). At baseline, HLA-DR was expressed in about 10 % of sputum T or B cells and a higher expression was found on monocytes. The HLA-DR expression on lymphocytes, but not monocytes, was significantly lower (p < 0.01) in patients treated with F + T. Fractions of activated [CD4⺠CD25âº] cells were also significantly lower in the combined therapy group, except for the subpopulation of CD4âºCD25(high) CD127(low) cells which was not altered. We conclude that tiotropium in add-on therapy to formoterol affects Treg cell profiles and decreases HLA-DR expression in airway lymphocytes.
Assuntos
Broncodilatadores/uso terapêutico , Etanolaminas/uso terapêutico , Antígenos HLA-DR/genética , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Derivados da Escopolamina/uso terapêutico , Linfócitos T Reguladores/efeitos dos fármacos , Agonistas de Receptores Adrenérgicos beta 2/uso terapêutico , Idoso , Antígenos CD/genética , Separação Celular , Antagonistas Colinérgicos/uso terapêutico , Quimioterapia Combinada , Fumarato de Formoterol , Expressão Gênica , Humanos , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Monócitos/patologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Escarro/citologia , Escarro/efeitos dos fármacos , Escarro/metabolismo , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/patologia , Brometo de TiotrópioRESUMO
Recent studies show that several Siglec receptors, such as Siglec-8 and Siglec-14, may be important therapeutic targets in asthma and COPD. Siglecs are a family of lectins belonging to the immunoglobulin superfamily and recognize sialic acid residues of glycoproteins. Most of Siglecs have intracellular immunoreceptor tyrosine-based inhibitory motifs (ITIM), implicating them in the suppression of immunoreceptor signaling. Siglec-5/14 may be involved in the negative regulation of innate immune responses. The aim of this study was to analyze Siglec-5/14 expression in induced sputum cells of COPD patients in the following treatment combinations: (1) a long-acting beta2-agonist, formoterol; (2) formoterol combined with a long-acting antimuscarinic agent, tiotropium; and (3) formoterol combined with an inhaled corticosteroid or formoterol combined with tiotropium and with an inhaled corticosteroid. Siglec expression was assessed in sputum cells by flow cytometry using a specific monoclonal antibody. Double staining of cells indicated that Siglec-5/14 is expressed in monocyte/macrophages and neutrophils, but not in lymphocytes. Siglec-5/14 expression was significantly higher in patients receiving combined therapy including inhaled corticosteroids compared with patients taking only formoterol or formoterol + tiotropium. Our results suggest that inhaled corticosteroids may exert beneficial or negative effects, depending on the patients' phenotype, through increased immunosuppressive Siglec-5 or immunoactivatory Siglec-14 receptors, respectively.
Assuntos
Corticosteroides/uso terapêutico , Antígenos CD/genética , Antígenos de Diferenciação Mielomonocítica/genética , Broncodilatadores/uso terapêutico , Etanolaminas/uso terapêutico , Lectinas/genética , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Receptores de Superfície Celular/genética , Derivados da Escopolamina/uso terapêutico , Agonistas de Receptores Adrenérgicos beta 2/uso terapêutico , Androstadienos/uso terapêutico , Budesonida/uso terapêutico , Separação Celular , Antagonistas Colinérgicos/uso terapêutico , Quimioterapia Combinada , Fluticasona , Fumarato de Formoterol , Expressão Gênica , Humanos , Imunofenotipagem , Lectinas/agonistas , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Neutrófilos/patologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Receptores de Superfície Celular/agonistas , Escarro/citologia , Escarro/efeitos dos fármacos , Escarro/metabolismo , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Linfócitos T/patologia , Brometo de TiotrópioRESUMO
The aim of this work was to assess the role of ethanol-derived acetate and acetate-mediated histone acetylation in arachidonic acid-induced stress in HepG2 cells and cells overexpressing CYP2E1. Cells were grown for 7 days with 1 mM sodium acetate or 100 mM ethanol; their acetylated histone proteins and histone deacetylase 2 expression was quantified using Western blot. Ethanol- or acetate-pretreated cells were also treated for 24 h with 60 µM arachidonic acid to induce oxidative stress. Cytotoxicity was estimated by lactate dehydrogenase release, 3-[4,5-dimethylthiazolyl-2] 2,5-diphenyltetrazolium bromide test, and by DNA damage, while oxidative stress was quantified using dichlorofluorescein diacetate. Cells grown with ethanol or acetate had increased acetylated histone H3 levels in both cell types and elevated acetylated histone H4 levels in cells overexpressing CYP2E1 but not in naïve cells. In cells overexpressing CYP2E1 grown with ethanol, expression of histone deacetylase 2 was reduced by about 40 %. Arachidonic acid altered cell proliferation and was cytotoxic mostly to cells engineered to overexpress CYP2E1 but both effects were significantly lower in cells pretreated with ethanol or acetate. Cytotoxicity was also significantly decreased by 4-methylpyrazole--a CYP2E1 inhibitor and by trichostatin--an inhibitor of histone deacetylases. In cells pretreated with acetate or ethanol, the oxidative stress induced by arachidonic acid was also significantly lower. Our data indicate that histone hyperacetylation may in some extent protect the cells against oxidative stress. It is possible that acetate may act as an antioxidant at histone level. This mechanism may be relevant to alcohol-induced liver injury.
Assuntos
Ácido Araquidônico/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Citocromo P-450 CYP2E1/metabolismo , Etanol/toxicidade , Acetatos/administração & dosagem , Acetatos/metabolismo , Acetilação , Antioxidantes/metabolismo , Etanol/metabolismo , Fomepizol , Células Hep G2 , Histonas/metabolismo , Humanos , Ácidos Hidroxâmicos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Pirazóis/farmacologiaRESUMO
Chronic obstructive pulmonary disease (COPD) is the only major disease with increasing death rate. In COPD, progressive reduction in quality of life is closely related to the increasing limitation of airflow due to chronic bronchitis, cell hyperplasia, fibrosis, and irreversible lung damage. Signaling pathways involved in inflammatory processes in COPD and inflammatory response to therapy are unknown. Our aim was to isolate cells from induced sputum of COPD patients treated with formoterol or formoterol + tiotropium and assess enzymatic activity of histone deacetylases (HDACs) acetylated histone 4 (AcH4) and expression of inducible nitric oxide synthase (iNOS). HDACs are important in signal transduction and inflammation. iNOS is generating nitric oxide (NO) relevant to blood pressure regulation, inflammation and infections. Thirty stable COPD patients (21 males and 9 females, mean age 67 years) receiving 12 µg b.i.d. formoterol were assayed before and after 3 months add-on therapy consisting of 18 µg q.i.d. tiotropium. In all patients, spirometry, lung volumes, and DLCO were performed before and after tiotropium therapy and all patients were subjected to sputum induction. Sputum cells were isolated and processed to obtain cytosolic and nuclear fractions. HDAC activity was measured in nuclear fraction using colorimetric assay. Expression AcH4 and iNOS was quantified using Western blot. In patients receiving both drugs, FEV1 and lung volumes significantly improved compared with formoterol-only treated patients. Mean HDAC activity was slightly decreased (P < 0.05), while AcH4 levels and iNOS expression were significantly elevated in tiotropium-treated patients (increase by about 65 %; P < 0.01 and 77 %; P < 0.01 respectively). Our data show that beneficial effects of tiotropium in add-on therapy to formoterol may be related to altered histone signaling and increased iNOS expression.
Assuntos
Regulação Enzimológica da Expressão Gênica , Histona Desacetilases/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Derivados da Escopolamina/farmacologia , Escarro/citologia , Idoso , Broncodilatadores/farmacologia , Núcleo Celular/metabolismo , Citosol/metabolismo , Etanolaminas/farmacologia , Feminino , Fumarato de Formoterol , Histonas/metabolismo , Humanos , Pulmão/enzimologia , Masculino , Óxido Nítrico/metabolismo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Transdução de Sinais , Fatores de Tempo , Brometo de TiotrópioRESUMO
Chronic obstructive pulmonary disease (COPD) is related to infiltration and activation of inflammatory cells in airways and pulmonary tissue. In COPD, neutrophils are prominent, while eosinophilic influx is typical to asthma. Inflammatory cells express sialic acid-binding immunoglobulin like lectins called Siglecs, a family of innate immune receptors that are transmembrane I-type lectins binding sialic acid. One member of the Siglec family, Siglec-8, is expressed mostly in eosinophils and may be an important therapeutic target in asthma or COPD. The aim of our project was to quantify Siglec-8 expression in induced sputum cells of COPD patients treated with long-acting beta2-agonists (LABA) or combined with long-acting antimuscarinic agents (LAMA) - tiotropium bromide. Thirty stable COPD patients (21 males and 9 females, mean age 67 years) receiving 12 µg BID formoterol therapy were assessed before and after 3 months' add-on therapy consisting of 18 µg QID tiotropium. In all patients, spirometry, lung volumes, and DLCO were performed before and after therapy. The patients were subjected to sputum induction before and after therapy. Sputum cells were isolated and processed to obtain cell membranes. Siglec-8 protein expression was assessed using Western blot. In patients receiving tiotropium and formoterol, improved FEV1 and lung volumes were observed compared with formoterol-only treated patients. The mean Siglec-8 level was significantly higher in eosinophilic subgroup of COPD patients compared with non-eosinophilic patients before therapy 40,000 vs. 15,000 Adj. Vol. INT/mm(2). Our data show that Siglec-8 may be involved in COPD pathogenesis and may influence COPD phenotyping.
Assuntos
Agonistas de Receptores Adrenérgicos beta 2/metabolismo , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos B/metabolismo , Regulação da Expressão Gênica , Lectinas/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Escarro/metabolismo , Idoso , Combinação de Medicamentos , Eosinófilos/metabolismo , Etanolaminas/administração & dosagem , Feminino , Fumarato de Formoterol , Humanos , Inflamação , Masculino , Fenótipo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Derivados da Escopolamina/administração & dosagem , Brometo de TiotrópioRESUMO
Chronic obstructive pulmonary disease (COPD) is characterized by progressive airflow limitation and chronic inflammation of airways and lung parenchyma. Our aim was to assess two important elements of intracellular signaling involved in regulation of inflammation in COPD in patients subjected to long-acting beta2-agonist or long-acting beta2-agonist plus long-acting antimuscarinic: peroxisome proliferator-activated receptor gamma (PPARγ) protein, which has antiinflammatory and immunomodulatory properties and cAMP response element binding protein (CREB) and activated (CREB-P) protein which has histone acetyltransferase activity and increases histone acetylation and transcriptional activation of chromatin. Twenty one stable COPD patients (18 males and 3 females, mean age 65 years) receiving 12 µg B.I.D formoterol were assayed before and after 3 month add-on therapy, consisting of 18 µg Q.D. tiotropium. In all patients, sputum induction, spirometry, lung volumes, and DLCO were performed before and after therapy. Sputum cells were isolated and processed to isolate cytosolic and nuclear fractions. PPARγ, CREB, or CREB-P proteins were quantified in subcellular fractions using Western blot. Tiotropium add-on therapy improved respiratory parameters: FEV1 and lung volumes. After therapy mean expression of PPARγ in cell nuclei was significantly increased by about 180%, while CREB and phosphorylated CREB levels in cytosol and nuclei were decreased by about 30%. Our data show that the mechanism whereby tiotropium reduces exacerbations may be associated not only with persistent increase in airway functions and reduced hyperinflation mediated by muscarinic receptors, but also with possible anti-inflammatory effects of the drug, involving increased PPARγ and decreased CREB signaling.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Antagonistas Muscarínicos/uso terapêutico , PPAR gama/metabolismo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Derivados da Escopolamina/uso terapêutico , Agonistas de Receptores Adrenérgicos beta 2/farmacologia , Agonistas de Receptores Adrenérgicos beta 2/uso terapêutico , Idoso , Anti-Inflamatórios não Esteroides/farmacologia , Broncodilatadores/farmacologia , Broncodilatadores/uso terapêutico , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Etanolaminas/farmacologia , Etanolaminas/uso terapêutico , Feminino , Volume Expiratório Forçado/efeitos dos fármacos , Fumarato de Formoterol , Humanos , Medidas de Volume Pulmonar , Masculino , Antagonistas Muscarínicos/farmacologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Testes de Função Respiratória , Derivados da Escopolamina/farmacologia , Transdução de Sinais , Escarro/química , Escarro/citologia , Brometo de TiotrópioRESUMO
Chronic obstructive pulmonary disease (COPD) is a progressive, inflammatory condition, involving airways and lung parenchyma. The disease leads to airflow limitation, and pulmonary hyperinflation, resulting in dyspnea, decreased exercise tolerance, and impaired quality of life. COPD pharmacotherapy guidelines are based on a combination of long-acting beta2-agonists (LABA), long-acting antimuscarinic agents (LAMA) and methyloxantins. Recently, indacaterol, ultralong acting beta2-agonist, has been introduced. The aim of our study was to assess the impact of indacaterol add-on therapy on lung function, exercise tolerance and quality of life of COPD patients. Thirty four COPD patients, receiving stable bronchodilator therapy were randomly allocated into two arms of add-on treatment (1:1 - indacaterol:placebo) for 3 months. Indacaterol replaced LABA in all patients receiving LABA. Spirometry, lung volumes, DLCO, St George's Respiratory Questionnaire (SGRQ) and 6 min Walk Distance (6MWD) were performed before and after therapy. We found that in the indacaterol group FEV1 did not changed significantly. However, there were significant improvements in ERV, 6MWD, and 6MWD-related dyspnea score. We also found that the degree of desaturation before and after 6MWD, and fatigue levels significantly improved in the indacaterol group. The patients' quality of life also changed favorably in the indacaterol treatment arm. We conclude that the add-on therapy with indacaterol exerts positive effects in COPD patients.
Assuntos
Tolerância ao Exercício/efeitos dos fármacos , Volume Expiratório Forçado/efeitos dos fármacos , Indanos/uso terapêutico , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Ventilação Pulmonar/efeitos dos fármacos , Quinolonas/uso terapêutico , Agonistas de Receptores Adrenérgicos beta 2 , Idoso , Broncodilatadores/uso terapêutico , Dispneia/tratamento farmacológico , Exercício Físico , Feminino , Humanos , Pulmão/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Antagonistas Muscarínicos/farmacologia , Antagonistas Muscarínicos/uso terapêutico , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Qualidade de Vida , Receptores Adrenérgicos beta 2/metabolismo , Testes de Função Respiratória , Inquéritos e QuestionáriosRESUMO
Angiogenesis plays important role in tumor growth and development. Protein ligands and their receptor tyrosine kinases are crucial in tumor related angiogenesis. Ligand/receptor systems such as vascular endothelial growth factor (VEGF), and tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains (Tie) family play important role in this phenomenon. The aim of this study was to evaluate the concentration of soluble receptor of VEGF (sVEGF R1) and Tie-2 domain in plasma of lung cancer patients before and after chemotherapy. Forty four lung cancer patients, 11 with small lung cancer (SCLC), 5 females and 6 males (mean age 60.2, range 39-72 years), and 33 patients with non-small cell lung cancer (N-SCLC), 6 females and 27 males (mean age 61.9, range 42-78 years) received four courses of chemotherapy. Control group consisted of 44 patients with COPD, 4 females and 40 males (mean age 37.1, 18-60 years). In all cases clinical partial response was achieved. Both sVEGF R1 and Tie-2 concentrations were elevated in cancer group before treatment compared with control: sVEGF (pg/ml): 60.7 and 66.2 vs. 48.8 and Tie-2 (ng/ml): 37.3 and 37.5 vs. 30.7 in SCLC and N-SCLC vs. C, respectively. Treatment decreased sVEGF R1 (pg/ml): 66.7 vs. 11.6 (p < 0.05) and 66.2 vs. 14.39 (p < 0.001), and Tie-2 (ng/ml): 37.3 vs. 26.3 (p < 0.05) and 37.5 vs. 25.7 (p < 0.001) in SCLC and N-SCLC, respectively. We conclude that VEGF R1and Tie-2 receptors may play important role in lung cancer development and their receptor concentrations may reflect the patients' response to treatment.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/tratamento farmacológico , Receptor TIE-2/sangue , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/sangue , Adulto , Idoso , Biomarcadores Tumorais , Cisplatino/uso terapêutico , Desoxicitidina/análogos & derivados , Desoxicitidina/uso terapêutico , Etoposídeo/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neovascularização Patológica , Doença Pulmonar Obstrutiva Crônica/sangue , Resultado do Tratamento , GencitabinaRESUMO
OBJECTIVE: Chronic obstructive pulmonary disease (COPD) is characterized by irreversible progressive airflow limitation related to tobacco smoking. This limitation is caused by chronic inflammation of the airways and lung parenchyma and is associated with increased activity of parasympathetic system. The most effective bronchodilators in COPD are muscarinic receptor antagonists (MRA), which reverse, at least in part, compromised respiratory function. MRA also contribute to control inflammatory processes via interactions with inflammatory signaling molecules. The use of the long-acting cholinolytic bronchodilatator - tiotropium, with high affinity to M3 receptors, is suggested as a first line maintenance treatment in COPD patients. MATERIAL AND METHODS: In this study we assessed M3 receptor protein expression in induced sputum of 27 stable COPD patients before and after therapy consisting of 18 µg once daily tiotropium for 12 weeks. Lung function tests including spirometry, lung volumes, and DLCO were performed before and after therapy in all COPD patients. The patients were subjected to the sputum induction procedures before and after therapy. Sputum cells were isolated, sample-specific cell profiles were characterized, and the cells were processed to isolate pure cytosolic fractions. Cytosolic M3 protein and HDAC2 levels and nuclear acetylated histone H3 (AcH3) expression was quantified using specific antibodies against human proteins and Western blot with enhanced luminescence detection. RESULTS: Therapy significantly increased the mean forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) volume (P<0.05). The mean expression of M3 protein was higher by 37% after therapy (P<0.05), HDAC2 expression was not altered, while AcH3 level was increased by about 90% (P<0.01), compared with the corresponding data before therapy. HDAC2 expression before therapy was positively correlated with AcH3 expression (r = 0.74), while after therapy no correlation was detected. FEV1, FCV, and cytosolic M3 protein expression did not correlate with other biochemical parameters tested. CONCLUSIONS: Twelve weeks of tiotropium therapy in COPD patients improves clinical indices of lung function and involves alterations in sputum cell chromatin acetylation and also increased cholinergic M3 receptor internalization.
Assuntos
Citosol/química , Histonas/metabolismo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Receptor Muscarínico M3/efeitos dos fármacos , Derivados da Escopolamina/farmacologia , Escarro/metabolismo , Acetilação , Volume Expiratório Forçado , Humanos , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Receptor Muscarínico M3/análise , Escarro/citologia , Brometo de Tiotrópio , Capacidade VitalRESUMO
OBJECTIVE: Heat shock proteins assist cellular protein folding and are required for the normal activity of steroid receptors. In this study we assessed nuclear HSP90 and HSP70 proteins and mRNA levels in cells isolated from induced sputum of chronic obstructive pulmonary disease patients treated for 4 weeks with formoterol (F) or formoterol+budesonide (F/ICS). METHODS: Nuclear heat shock protein levels were assessed by Western blot and specific mRNAs were quantified in cell lysates using qRT-PCR. RESULTS: Both HSP90 and HSP70 protein levels were higher in the F/ICS-treated patients in comparison with the F-treated group (by 31%, P<0.05 and 28%, P<0.05, respectively), while specific mRNAs were lowered. HSP86/HSP89 and D6S182/HSP90-BETA were repressed by about 40% (P<0.05) while HSP70-1/HSP70-1A, HSP70-1B/HSP70-2, and HSP70-HSC54/HSC70 were repressed by 47% (P<0.01), 57% (P<0.01) and 65% (P<0.01), respectively. CONCLUSIONS: It is possible that increased nuclear heat shock proteins may play a role in the attenuation of the response to glucocorticoids in COPD patients.
Assuntos
Budesonida/administração & dosagem , Núcleo Celular/química , Etanolaminas/uso terapêutico , Proteínas de Choque Térmico HSP70/análise , Proteínas de Choque Térmico HSP90/análise , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Quimioterapia Combinada , Etanolaminas/administração & dosagem , Fumarato de Formoterol , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP90/genética , Humanos , Doença Pulmonar Obstrutiva Crônica/metabolismo , RNA Mensageiro/análiseRESUMO
OBJECTIVE: Immunophilin FKBP51 assists polypeptide folding, participates in glucocorticoid actions and may play a role in glucocorticoid resistance. FKBP51 is altered in patients with asthma, but its role in chronic obstructive pulmonary disease (COPD) characterized by dysregulation of several pro/antiinflammatory genes is less clear. METHODS: We assessed changes in nuclear/cytosolic FKBP51 protein using SDS-PAGE/WB and FKBP51 mRNA by qRT-PCR in cells isolated from induced sputum of stable COPD patients treated with formoterol/budesonide or formoterol/budesonide/theo?phylline for 4 wk. RESULTS: Expression of FKBP51 was higher in formoterol/ budesonide/theophylline-treated patients, compared with formoterol/budesonide group in both cytosolic and nuclear fractions by about 57% and 31%, respectively (P<0.001, P<0.01). FKBP51 mRNA was only slightly, but not significantly, higher in patients on formoterol/ budesonide/theophylline. CONCLUSIONS: Increased FKBP51 in COPD patients treated with formoterol/ budesonide/theophylline may be important in altering signaling from corticosteroid receptors.
Assuntos
Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Escarro/química , Proteínas de Ligação a Tacrolimo/análise , Budesonida/administração & dosagem , Quimioterapia Combinada , Etanolaminas/administração & dosagem , Fumarato de Formoterol , Humanos , Doença Pulmonar Obstrutiva Crônica/metabolismo , RNA Mensageiro/análise , Proteínas de Ligação a Tacrolimo/genética , Teofilina/administração & dosagemRESUMO
A 61-year-old Caucasian was admitted to Department of Chest Diseases and Tuberculosis, Medical University of Bialystok, Poland for progressive muscle weakness and weight loss. Eighteen months prior to admission, the patient had been diagnosed with pulmonary embolism. At that point he was started on Enoxaparin QD. Past medical history was unremarkable. In the interim, the patient developed fever, myalgia and progressive dyspnea. Physical examination on admission revealed a rash on his upper torso and back, and the extensor surfaces of all four extremities. Laboratory values included CPK 8229, MB fraction 219, LDH 981. Chest X-ray and CT scan revealed bilateral patchy consolidations and ground-glass opacities. EMG was consistent with myositis. The patient was started on solumedrol 40 mg i.v., b.i.d., and then switched to prednisone 40 mg b.i.d. His symptoms and muscle strength improved remarkably. The patient was discharged with prednisone with an outpatient follow up.
Assuntos
Dermatomiosite/diagnóstico , Embolia Pulmonar/diagnóstico , Dermatomiosite/tratamento farmacológico , Dermatomiosite/etiologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: It has recently been described that circulatory and BAL regulatory T-cells (Tregs), defined as CD4+CD25highCD127low are increased in patients with active sarcoidosis compared with other interstitial lung diseases. MATERIAL AND METHODS: We studied prospectively 17 patients (10 women, 7 men) of median age 39 years (range 27-65) with active granulomatous lung diseases (GLD) (10 patients with sarcoidosis (BBS), and 7 with hypersensitivity pneumonitis (HP), and 9 healthy controls. Bronchoalveolar lavage fluid (BAL) and induced sputum Treg counts, CD4+, CD8+, CD25+ cells were quantified by flow cytometry. Disease activity was measured by ACE serum level. Pulmonary function tests were performed using an Elite DL Medgraphics body box. RESULTS: We found Treg cells count significantly elevated in induced sputum from active GLD (38.3% vs. 7.1% and 5.3% in BBS, HP, and control, respectively). A significantly higher percentage of Treg cells characterized BAL cells from HP patients (2.27%; 9.5%; 2.1%, in BBS, HP and control, respectively). There was a strong correlation with ACE serum level and Treg cell count in BAL fluid of BBS patients, with no such correlation within HP patient group, nor Treg cell count and pulmonary function tests. CONCLUSIONS: Our data suggest a potential role of CD4+CD25highCD127low induced sputum and BAL lymphocytes from patients with active granulomatous lung diseases and hypersensitivity pneumonitis. An increased number of Treg cells in active GLD may be involved in immune regulation in active granulomatous lung diseases. The results indicate that analysis of these cells could be useful as markers of disease activity in granulomatous lung diseases.
Assuntos
Líquido da Lavagem Broncoalveolar/imunologia , Sarcoidose Pulmonar/imunologia , Escarro/imunologia , Adulto , Idoso , Relação CD4-CD8 , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Linfócitos T ReguladoresRESUMO
Propofol can be potentially beneficial in oxidative stress related malignancies as neurodegenerative diseases and traumatic brain injury but its signalling pathways are poorly understood. In this study effect of propofol on astroglial signalling in oxidative stress was evaluated. Ten days old cultures of rat astroglial cells were treated for 1 hour with t-butyl hydroperoxide (tBHP) to induce oxidative stress following by 1 hour propofol. We measured cytotoxicity, changes in cell growth and apoptosis as well as alterations in expression and acetylation of chromatin core H3 and H4 histone proteins and changes in native and phosphorylated cAMP-response-element-binding protein (CREB). tBHP induced limited cytotoxicity, increased apoptosis, decreased glutamine synthetase and enolase activities, decreased nuclear CREB, CREB-P and histone proteins but unchanged cytosolic CREB and histone acetyltransferase (HDAC) expression. Propofol clearly protected the cells against tBPH-induced toxicity, normalized alterations in cell growth, restored to some extent glial enzyme activities and reduced apoptotic cell numbers. Also, propofol restored H3 but not H4 expression/activation, but was without effect on decreased nuclear CREB expression/activation. These data show that oxidative stress in cultured astroglia significantly affects nuclear CREB and histone proteins and point to the protective role of propofol.
Assuntos
Anestésicos Intravenosos/farmacologia , Astrócitos/efeitos dos fármacos , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Histonas/metabolismo , Propofol/farmacologia , Transdução de Sinais/efeitos dos fármacos , terc-Butil Hidroperóxido/toxicidade , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cromatina/efeitos dos fármacos , Glutamato-Amônia Ligase/metabolismo , Histona Acetiltransferases/metabolismo , Histonas/isolamento & purificação , Estresse Oxidativo/efeitos dos fármacos , Fosfopiruvato Hidratase/metabolismo , Fosforilação/efeitos dos fármacos , Transporte Proteico/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
The epidemiological association between cancer and exposure to ambient air pollution particles (particles with a 50% cut-off aerodynamic diameter of 10 microm (PM(10))) has been related to the ability of PM(10) and its constituent nanoparticles (NPs) to cause reactive oxidative species (ROS)-driven DNA damage. However, there are no data on the molecular response to these genotoxic effects. In order to assess whether PM(10), NP and ROS-driven DNA damage induce carcinogenesis pathways, A549 cells were treated with tert-butyl-hyperperoxide (Tbh), urban dust (UD), carbon black (CB), nanoparticulate CB (NPCB), benzo(a)pyrene (BaP) and NPCB coated with BaP for Assuntos
Carcinógenos Ambientais/toxicidade
, Transformação Celular Neoplásica/induzido quimicamente
, Dano ao DNA/efeitos da radiação
, Nanopartículas/toxicidade
, Análise de Variância
, Western Blotting
, Linhagem Celular Tumoral
, Ensaio Cometa
, Quebras de DNA de Cadeia Dupla
, Genes BRCA1
, Humanos
, Técnicas In Vitro
, Análise Multivariada
, Mutagênicos/toxicidade
, Estresse Oxidativo
, Fosforilação
, Espécies Reativas de Oxigênio/metabolismo
, Sensibilidade e Especificidade
RESUMO
Chronic obstructive pulmonary disease (COPD) is one of the most frequent diseases worldwide. Cigarette smoke is considered the main pathological cause of the disorder, although evidence is growing concerning other etiological factors, such as environmental pollution, biomass combustion, infections, genetic predisposition, which may explain why some individuals develop COPD with no history of smoking. Chronic inflammation and remodeling of the small airways characterize the disease at the cellular level, and oxidative stress is considered the main driving force that stands behind COPD inflammation. Recently, chromatin remodeling and epigenetic changes have been found to underlie disease pathology and progression. In this review, the authors gave a short update on the recent hypothesis and findings that may imply novel approach to pharmacotherapy of the disease, focusing on the role of glucocorticosteroids, theophylline, and antioxidants.
Assuntos
Anti-Inflamatórios/uso terapêutico , Núcleo Celular/patologia , Inflamação/patologia , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/patologia , Transdução de Sinais/efeitos dos fármacos , Animais , Cromatina/efeitos dos fármacos , Cromatina/ultraestrutura , Glucocorticoides/farmacologia , Glucocorticoides/fisiologia , Glucocorticoides/uso terapêutico , Humanos , Doença Pulmonar Obstrutiva Crônica/genética , Receptores de Glucocorticoides/fisiologia , Fatores de Transcrição/fisiologiaRESUMO
We assessed the effect of therapy on nuclear signaling related to inflammatory processes in sputum cells of patients with chronic obstructive pulmonary disease (COPD). Patients were treated with formoterol (F) or formoterol plus budesonide (F/ICS) b.i.d. for 4 weeks, their sputum cells were isolated and subjected to RNA extraction or lysis, followed by differential centrifugation. Signaling protein levels were assessed by Western blots, their specific mRNAs were quantified using qRTPCR, while 8-isoprostane levels were examined using enzyme immunoassay kit. Cytosolic 8-isoprostane levels and nuclear glucocorticoid receptor expression (protein and mRNA) were not significantly different in both groups, while nuclear cAMP response element binding protein (CREB; protein and mRNA) and peroxisome proliferator-activated receptor gamma (PPARgamma protein and mRNA) were significantly higher in cells from F/ICS-treated patients. CREB-binding protein (CBP; protein and mRNA) levels were significantly lower in F/ICS patients. These changes indicate increased anti-inflammatory signaling in F/ICS-treated patients and seem to be beneficial.
Assuntos
Broncodilatadores/uso terapêutico , Proteína de Ligação a CREB/biossíntese , Etanolaminas/uso terapêutico , Glucocorticoides/uso terapêutico , PPAR gama/biossíntese , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Western Blotting , Dinoprosta/análogos & derivados , Dinoprosta/sangue , Quimioterapia Combinada , Ensaio de Imunoadsorção Enzimática , Volume Expiratório Forçado/fisiologia , Fumarato de Formoterol , Humanos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Doença Pulmonar Obstrutiva Crônica/complicações , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Receptores de Glucocorticoides/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Escarro/citologiaRESUMO
We investigate retrospectively the efficacy of self-expandable metallic stents (SEMS) for severe respiratory distress (SRD) in patients with central airway obstruction (CAO). Thirty three patients with CAO were treated with SEMS using fiberoptic bronchoscopy method. We found an intraluminal obstruction present in 7, extraluminal compression in 10, and combined stenosis in 16 cases. Tumor infiltration occupied more than 90% of the endoluminal diameter in 21, 70% in 9, and 50% in 3 cases. Obstruction was caused by primary cancer of lung in 23, thyroid in 5, and esophagus in 5 patients. Up to 3 stents per patient were placed. Double stenting (esophagus and trachea) was required in five patients. All patients exhibited symptomatic and arterial blood gas improvement. The mean follow-up was 65 (5 to 752) days. SEMS are useful for the treatment of SRD caused by CAO. The overall effect is related to the degree of tumor progression itself.