Asthma and lower airway diseaseSteroids completely reverse albuterol-induced β2-adrenergic receptor tolerance in human small airways
Section snippets
Reagents
Reagents used were carbachol, isoproterenol, low-melting-point agarose (IX-A), dexamethasone, and Ham's F-12 medium (supplemented with 2 mmol/L glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin, 2.5 μg/mL Fungizone, 50 μg/mL gentamycin, and 1 mol/L HEPES [pH 7.6]). All reagents were obtained from Sigma (St Louis, Mo), unless otherwise stated.
PCLS preparation and airway function
Healthy whole lungs were received from the National Disease Research Interchange. The smallest lobe was cut free, exposing its main bronchiole, and
Carbachol induces airway luminal diameter narrowing in a dose-dependent manner
Slices were incubated with cumulative doses of carbachol and luminal narrowing was determined as shown in Fig 1, A, to determine whether agonists induce small airway narrowing in PCLSs. Carbachol abrogated airway luminal diameter with a log EC50 value of −0.39 ± 0.06 μmol/L. Additionally, the Emax (86.0% ± 3.1%) of this effect was observed at a concentration of 30 μmol/L. Carbachol dose responses inducing luminal diameter narrowing were performed at 24, 48, 72, and 96 hours ex vivo to assess
Discussion
This study is the first to demonstrate a model of β2-AR tolerance in human small airways. The short-acting bronchodilator albuterol induced a concentration- and time-dependent decrease in β2-AR activity at 3, 6, and 12 hours and at concentrations of 0.01, 0.1, and 1.0 μmol/L, with no difference in relaxation to forskolin shown after incubation with albuterol versus control values. This study has also shown that a 1-hour incubation with dexamethasone prevented the β2-AR desensitization from
References (18)
- et al.
Rapid onset of tolerance to beta-agonist bronchodilation
Respir Med
(2005) - et al.
Measurement of protein using bicinchoninic acid
Anal Biochem
(1985) - et al.
Spontaneous and provoked resistance to isoproterenol in isolated human bronchi
J Allergy Clin Immunol
(1984) - et al.
An in vivo model of beta-adrenoceptor desensitization
J Pharmacol Toxicol Methods
(1998) - et al.
A bolus of inhaled budesonide rapidly reverses airway subsensitivity and beta2-adrenoceptor down-regulation after regular inhaled formoterol
Chest
(1999) - et al.
The synthesis of beta-adrenergic receptors in cultured human lung cells: induction by glucocorticoids
Biochem Biophys Res Commun
(1980) - et al.
Long-term effects of a long-acting beta 2-adrenoceptor agonist, salmeterol, on airway hyperresponsiveness in patients with mild asthma
N Engl J Med
(1992) - et al.
Effect of inhaled formoterol and budesonide on exacerbations of asthma. Formoterol and Corticosteroids Establishing Therapy (FACET) International Study Group
N Engl J Med
(1997) - et al.
Long-acting beta-agonist treatment in patients with persistent asthma already receiving inhaled corticosteroids
BioDrugs
(2001)
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2020, Pharmacology and TherapeuticsCitation Excerpt :This GC effect may be even more pronounced under conditions in which the β2 adrenergic receptor/Gs/AC/cAMP/PKA pathway is impaired due to β2-receptor desensitization, a feature that may develop in patients with severe asthma (Chachi et al., 2018) or after long-term treatment with β2-agonists (Amrani & Bradding, 2017). Although GCs prevent β2-receptor desensitization in some model systems, including precision-cut lung slices (Cooper & Panettieri Jr., 2008), the clinical relevance of such in vitro observations remains to be confirmed since poor responses to β2-receptor agonists is a key feature of patients with severe asthma despite being treated with high doses of inhaled or oral GCs (Chachi et al., 2018). As discussed previously, the efficacy of GCs in suppressing the expression of pro-inflammatory genes is greatly influenced by the type of stimulus and associated signaling pathways.
Supported by HL080676, HL064063, HL081824, and ES013508.
Disclosure of potential conflict of interest: The authors have declared that they have no conflict of interest.