BACKGROUND: Neutrophils and oxidative stress have been implicated in the pathogenesis of COPD. Severe, early-onset COPD is characterized by a rapid decline in the lung function at an early age; however, nothing is known about neutrophil activation in COPD patients. OBJECTIVES: The aim of this study was to evaluate peripheral blood neutrophil activation in severe, early-onset COPD patients versus healthy non-smokers and the effect of N-acetyl-L-cysteine (NAC) on neutrophil activation in vitro. METHODS: Neutrophils were isolated from 15 severe, early-onset COPD patients and 15 age-matched healthy subjects and stimulated with N-formyl-Met-Leu-Phe (fMLP) in the presence or absence of NAC (10 μM to 10 mM). Neutrophil chemotaxis, elastase release, reactive oxygen species (ROS), intracellular thiols and apoptosis were measured by Boyden chamber, spectrofluorometry, CMFDA and H2DCF-DA dyes and by annexin V-FITC, respectively. RESULTS: Chemotaxis of peripheral blood neutrophils from COPD patients in response to fMLP was 30% more increased than that observed in healthy subjects. Elastase release in response to fMLP was 2-fold higher in neutrophils from COPD patients versus healthy subjects. Intracellular thiol levels were 30% lower in COPD and ROS was approximately 30% higher in COPD versus healthy neutrophils. Spontaneous apoptosis showed no differences in both groups of patients and fMLP-induced apoptosis was higher in COPD. Pre-treatment with the antioxidant NAC effectively attenuated neutrophil chemotaxis, elastase release and ROS as well as effectively increased thiol levels in COPD. CONCLUSIONS: Neutrophils in severe, early-onset COPD patients are highly activated and this is alleviated by NAC in vitro.
BACKGROUND: Neutrophils and oxidative stress have been implicated in the pathogenesis of COPD. Severe, early-onset COPD is characterized by a rapid decline in the lung function at an early age; however, nothing is known about neutrophil activation in COPDpatients. OBJECTIVES: The aim of this study was to evaluate peripheral blood neutrophil activation in severe, early-onset COPDpatients versus healthy non-smokers and the effect of N-acetyl-L-cysteine (NAC) on neutrophil activation in vitro. METHODS: Neutrophils were isolated from 15 severe, early-onset COPDpatients and 15 age-matched healthy subjects and stimulated with N-formyl-Met-Leu-Phe (fMLP) in the presence or absence of NAC (10 μM to 10 mM). Neutrophil chemotaxis, elastase release, reactive oxygen species (ROS), intracellular thiols and apoptosis were measured by Boyden chamber, spectrofluorometry, CMFDA and H2DCF-DA dyes and by annexin V-FITC, respectively. RESULTS: Chemotaxis of peripheral blood neutrophils from COPDpatients in response to fMLP was 30% more increased than that observed in healthy subjects. Elastase release in response to fMLP was 2-fold higher in neutrophils from COPDpatients versus healthy subjects. Intracellular thiol levels were 30% lower in COPD and ROS was approximately 30% higher in COPD versus healthy neutrophils. Spontaneous apoptosis showed no differences in both groups of patients and fMLP-induced apoptosis was higher in COPD. Pre-treatment with the antioxidant NAC effectively attenuated neutrophil chemotaxis, elastase release and ROS as well as effectively increased thiol levels in COPD. CONCLUSIONS: Neutrophils in severe, early-onset COPDpatients are highly activated and this is alleviated by NAC in vitro.
Authors: A Brito; T Santos; K Herculano; M Miranda; A K Sá; J L Carvalho; R Albertini; H Castro-Faria-Neto; A P Ligeiro-de-Oliveira; Flávio Aimbire Journal: Inflammation Date: 2021-03-17 Impact factor: 4.092
Authors: Tina Kirchner; Eva Hermann; Sonja Möller; Matthias Klinger; Werner Solbach; Tamás Laskay; Martina Behnen Journal: Mediators Inflamm Date: 2013-12-07 Impact factor: 4.711
Authors: Alexandra Sakelliou; Ioannis G Fatouros; Ioannis Athanailidis; Dimitrios Tsoukas; Athanasios Chatzinikolaou; Dimitris Draganidis; Athanasios Z Jamurtas; Christina Liacos; Ioannis Papassotiriou; Dimitrios Mandalidis; Kimon Stamatelopoulos; Meletios A Dimopoulos; Asimina Mitrakou Journal: Oxid Med Cell Longev Date: 2016-11-15 Impact factor: 6.543
Authors: Mike Lonergan; Alison J Dicker; Megan L Crichton; Holly R Keir; Melissa K Van Dyke; Hana Mullerova; Bruce E Miller; Ruth Tal-Singer; James D Chalmers Journal: Respir Res Date: 2020-07-01