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1
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Differences in responses upon corticosteroid therapy between smoking and non-smoking patients with COPD

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Van Overveld F. J., Demkow U., Gorecka D., De Backer W. A., Zielinski J.
Journal of Physiology and Pharmacology. Supplement
|
2006
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tom 57
|
nr 4
273-282
Inhaled corticosteroids have a high level of topical anti-inflammatory activity. However, in patients with COPD these drugs have been reported to exert limited effects. A reduction in histone deacetylase (HDAC) activity is suggested to prevent the anti-inflammatory action of corticosteroids. Cigarette smoke is known to reduce HDAC expression. The aim of this study is to compare the outcome of corticosteroid therapy in both smoking and non-smoking COPD patients. Twenty-three smoking patients and 18 ex-smoking patients with COPD were treated with inhaled corticosteroids for a period of 2 months. Blood and induced sputum samples were collected before and after treatment. Values of FEV1 %-predicted did not change upon the therapy, but there was a trend to improve in the ex-smokers (63.1 64.8%-pred.), compared with a decrease in the smokers (63.3 61.6%-pred.). The levels of the pro-inflammatory cytokine IL-8 increased in the group of smokers from 379 ±78 to 526 ±118 ng/ml. Although not significant, a slight decrease from 382 ±70 to 342 ±62 ng/ml was observed in the group of ex-smokers. The neutrophil related elastase activity showed similar effects after steroid treatment, it went up from 36.4 ±12.0 to 113.5 ±9.7 nmol/l in smokers, and decreased from 346.2 ±72.1 to 131.1 ±6.5 nmol/l in ex-smokers with COPD. These results support the evidence that inhaled corticosteroids have no anti-inflammatory effects in COPD patients, but only when these patients are still smoking. Smoking cessation seems the best therapy for COPD patients.
2
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New developments in the treatment of COPD: comparing the effects of inhaled corticosteroids and N-acetylcysteine

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Van Overveld F. J., Demkow U., Gorecka D., De Backer W. A., Zielinski J.
Journal of Physiology and Pharmacology. Supplement
|
2005
|
tom 56
|
nr 4
135-142
Inhaled corticosteroids (ICS) are widely used for the treatment of COPD despite of controversial statements concerning their efficacy. The use of N-acetylcysteine (NAC), a mucolytic drug with antioxidant properties, is less clear, but it may counteract the oxidant-antioxidant imbalance in COPD. The aim of this study was to evaluate whether treatment of COPD patients with ICS or NAC is able to improve inflammatory indices and to enhance lung function. ICS treatment enhanced protective markers for oxidative stress such as glutathione peroxidase (GPx) (51.2 ±5.8 vs. 62.2 ±8.6 U/g Hb, P<0.02) and trolox-equivalent antioxidant capacity (TEAC) (1.44 ±0.05 vs. 1.52 ±0.06 mM, P<0.05). NAC decreased sputum eosinophil cationic protein (318 ±73 vs. 163 ±30 ng/ml, P<0.01) and sputum IL-8 (429 ±80 vs. 347 ±70 ng/ml, P<0.05). The increased antioxidant capacity prevented an up-regulation of adhesion molecules, since the levels of intracellular adhesion molecule 1 (ICAM-1) correlated negatively with GPx (P<0.0001) and TEAC (P<0.0001). On the other hand, expression of adhesion molecules was promoted by inflammation, reflected by a positive correlation between the levels of IL-8 and ICAM-1 (P<0.0001). The effects of treatment on lung function were only reflected in the FEV1 values. The absolute value of FEV1, both before and after salbutamol inhalation, increased from 1690 ±98 to 1764 ±110 ml, and 1818 ±106 to 1906 ±116 ml, respectively, after ICS (P<0.05) . Ten weeks after treatment, FEV1 values dropped to 1716 ±120 ml post-salbutamol (P<0.05). When followed by treatment with NAC, these values decreased even further to 1666 ±84 ml. These results suggest that ICS improved lung function in COPD patients with moderate airflow obstruction, beside a minor improvement in the oxidant-antioxidant imbalance leading to a lesser expression of ICAM-1. Treatment with NAC decreased some inflammatory parameters and had indirectly an inhibitory effect on the expression of adhesion molecules.
3
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Effects of homogenization of induced sputum by dithiothreitol on polymorphonuclear cells

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Van Overveld F. J., Demkow U., Gorecka D., Skopinska-Rozewska E., De Backer W. A., Zielinski J.
Journal of Physiology and Pharmacology. Supplement
|
2005
|
tom 56
|
nr 4
143-154
Induced sputum represents a useful and non-invasive tool to isolate different cells from the airways. Complete homogenization of sputum is important for dispersion of cells and is usually achieved by use of dithiothreitol (DTT). However, it is not known if DTT will influence the viability and functionality of cells obtained by induced sputum. In the present study, induced sputum was processed by DTT or by PBS treatment. The obtained neutrophils were compared with neutrophils obtained from peripheral blood and from bronchoalveolar lavage fluid (BAL). These isolated neutrophils were treated in a similar way as the sputum neutrophils with DTT or PBS. All isolated cells were used for chemiluminescence tests and for the measurement of elastase and myeloperoxidase release after stimulation with fMLP. The results showed that the maximum chemiluminescence response was always significantly lower after DTT treatment: blood, 16.68 ±1.89 vs. 2.62 ±0.43 mV, P<0.0001; sputum, 2.96 ±0.30 vs. 1.09 ±0.01 mV, P<0.01; BAL, 25.47 ±0.88 vs. 8.22±0.20 mV, P<0.0001. Both spontaneous and fMLP-induced release of elastase and myeloperoxidase (MPO) was in most cases enhanced after DTT-treatment (P-values range from 0.24 to <0.01). We conclude that the use of DTT to homogenize sputum for dispersion of cells is harmful to cell functions and these cells are hampered for the evaluation of their normal functional characteristics.
4
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Influence of N-acetylcysteine on ICAM-1 expression and IL-8 release from endothelial and epithelial cells

86%
Radomska-Lesniewska D. M., Sadowska A. M., Van Overveld F. J., Demkow U., Zielinski J., De Backer W. A.
Journal of Physiology and Pharmacology. Supplement
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2006
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tom 57
|
nr 4
325-334
Chronic obstructive pulmonary disease (COPD) is characterized by chronic airway inflammation. The initial step in the inflammatory process is overexpression of adhesion molecules, which leads to excessive transmigration of neutrophils. One of these adhesion molecules is ICAM-1 which is elevated in COPD patients. In this study we evaluated the influence of N-acetylcysteine (NAC) (0.01 mM-30 mM) on the cytokine-induced (TNF-alphalpha/IL-1ß) expression of the ICAM-1 adhesion molecule and on IL-8 release in endothelial (ECV-304) and bronchial epithelial (H292) cell lines. The methodology used consisted of immunochemistry for the assessment of surface ICAM-1 and ELISA method for that of soluble ICAM-1 and IL-8. NAC inhibited the TNF-alphalpha/IL-1ß-stimulated ICAM-1 expression and IL-8 release from both cell lines in a concentration dependent manner. The most effective concentrations were 30 mM and 20 mM (99 and 90% inhibition respectively, P<0.01). We conclude that NAC is an effective inhibitor of TNF-alphalpha/IL-1ß- stimulated ICAM-1 and IL-8 release in endothelial and epithelial cells. This fact highlights the anti-inflammatory potential of NAC in COPD.
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