Non-neuronal cholinergic system contributes to corticosteroid resistance in chronic obstructive pulmonary disease patients.
Inhaled corticosteroid (ICS) with long-acting beta-2 agonists is a well-documented combination therapy for chronic obstructive pulmonary disease (COPD) based on its additive anti-inflammatory properties. By contrast, the recommendation of ICS in combination with long-acting muscarinic antagonist (LAMA) is not evidence-based. In this study, neutrophils obtained from COPD patients were used to compare the anti-inflammatory effects of aclidinium bromide (a long-acting muscarinic antagonist) with corticosteroids and their potential additive effect.



Human sputum and blood neutrophils were isolated from healthy individuals (n = 37), patients with stable COPD (n = 52) and those with exacerbated COPD (n = 16). The cells were incubated with corticosteroid fluticasone propionate (0.1 nM-1 μM), aclidinium bromide (0.1 nM-1 μM) or a combination thereof and stimulated with 1 μg of lipopolysaccharide/ml or 5 % cigarette smoke extract. Levels of the pro-inflammatory mediators interleukin-8, matrix metalloproteinase-9, CCL-5, granulocyte-macrophage colony-stimulating factor and interleukin-1β were measured and the mechanisms of corticosteroid resistance evaluated at the end of the incubation.



The non-neuronal cholinergic system was over-expressed in neutrophils from COPD patients, as evidenced by increases in the expression of muscarinic receptors (M2, M4 and M5), choline acetyltransferase and vesicular acetylcholine transporter. Aclidinium bromide demonstrated anti-inflammatory effects on neutrophils from COPD patients, reversing their resistance to corticosteroids. Additive effects of combined aclidinium bromide and fluticasone propionate in blocking M2 receptor levels, inhibiting phosphoinositide 3-kinase-δ and enhancing the glucocorticoid response element transcription factor were demonstrated and were accompanied by an increase in the corticosteroid-induced expression of anti-inflammatory-related genes.



LAMAs potentiate the anti-inflammatory effects of corticosteroids in neutrophils from COPD patients in vitro, thus providing a scientific rationale for their use in combination with corticosteroids in the treatment of COPD.
Department of pharmacology, faculty of medicine, Jaume I University, Castellón, Spain. xmilara@hotmail.com.|Pharmacy Unit, University General Hospital Consortium, Valencia, Spain. xmilara@hotmail.com.|CIBERES, Health Institute Carlos III, Valencia, Spain. xmilara@hotmail.com.|Unidad de Investigación Clínica, Consorcio Hospital General Universitario, Avenida tres cruces s/n, E-46014, Valencia, Spain. xmilara@hotmail.com.
Javier
J
Milara
Respiratory Unit, University General Hospital Consortium, Valencia, Spain.
Angela
A
Cervera
Respiratory Unit, University and Polytechnic La Fe Hospital, Valencia, Spain.
Alfredo
A
de Diego
Department of pharmacology, faculty of medicine, Jaume I University, Castellón, Spain.|Department of Pharmacology, Faculty of Medicine, University of Valencia, Valencia, Spain.
Celia
C
Sanz
Respiratory Unit, University General Hospital Consortium, Valencia, Spain.
Gustavo
G
Juan
Almirall, R&D Centre, Barcelona, Spain.
Amadeu
A
Gavaldà
Almirall, R&D Centre, Barcelona, Spain.
Montserrat
M
Miralpeix
CIBERES, Health Institute Carlos III, Valencia, Spain.|Department of Pharmacology, Faculty of Medicine, University of Valencia, Valencia, Spain.|Health Research Institute INCLIVA, Valencia, Spain.
Esteban
E
Morcillo
CIBERES, Health Institute Carlos III, Valencia, Spain.|Department of Pharmacology, Faculty of Medicine, University of Valencia, Valencia, Spain.|Research and teaching Unit, University General Hospital Consortium, Valencia, Spain.
Julio
J
Cortijo
D000368:Aged; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001993:Bronchodilator Agents / Q000494:pharmacology*; D016022:Case-Control Studies; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance / Q000187:drug effects*; D004357:Drug Synergism; D004359:Drug Therapy, Combination; D005260:Female; D000068298:Fluticasone / Q000494:pharmacology*; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D018727:Muscarinic Antagonists / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D000070634:Non-Neuronal Cholinergic System / Q000187:drug effects*; D029424:Pulmonary Disease, Chronic Obstructive / Q000097:blood / Q000188:drug therapy*; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism; D013183:Sputum / Q000378:metabolism; D014326:Tropanes / Q000494:pharmacology*; D050494:Vesicular Acetylcholine Transport Proteins / Q000378:metabolism
D000893:Anti-Inflammatory Agents; D001993:Bronchodilator Agents; D018836:Inflammation Mediators; D018727:Muscarinic Antagonists; D011976:Receptors, Muscarinic; D014326:Tropanes; D050494:Vesicular Acetylcholine Transport Proteins; D000068298:Fluticasone; D002795:Choline O-Acetyltransferase; C542859:aclidinium bromide
Aclidinium bromide; COPD; Corticosteroid resistance; Neutrophils; Non-neuronal cholinergic system