Research

Cysteinyl-leukotrienes in the regulation of β₂-adrenoceptor function: an in vitro model of asthma

G Enrico Rovati¹ , Michele Baroffio^2* , Simona Citro^1* , Lorenzo Brichetto² , Saula Ravasi¹ , Manlio Milanese² , Emanuele Crimi² and Vito Brusasco²

¹  Laboratory of Molecular Pharmacology, Section of Eicosanoid Pharmacology, Dept. of Pharmacological Sciences, University of Milan, Italy

²  Respiratory Pathophysiology Unit, Dept. of Internal Medicine, University of Genoa, Italy

author email corresponding author email* Contributed equally

Respiratory Research 2006, 7:103doi:10.1186/1465-9921-7-103

Published:	28 July 2006

Abstract

Background

The response to β₂-adrenoceptor agonists is reduced in asthmatic airways. This desensitization may be in part due to inflammatory mediators and may involve cysteinyl-leukotrienes (cysteinyl-LTs). Cysteinyl-LTs are pivotal inflammatory mediators that play important roles in the pathophysiology of asthma, allergic rhinitis, and other inflammatory conditions. We tested the hypothesis that leukotriene D₄ (LTD₄) and allergen challenge cause β₂-adrenoceptor desensitization through the activation of protein kinase C (PKC).

Methods

The isoproterenol-induced cAMP accumulation was evaluated in human airway smooth muscle cell cultures challenged with exogenous LTD₄ or the PKC activator phorbol-12-myristate-13-acetate with or without pretreatments with the PKC inhibitor GF109203X or the CysLT₁R antagonist montelukast. The relaxant response to salbutamol was studied in passively sensitized human bronchial rings challenged with allergen in physiological salt solution (PSS) alone, or in the presence of either montelukast or GF109203X.

Results

In cell cultures, both LTD₄ and phorbol-12-myristate-13-acetate caused significant reductions of maximal isoproterenol-induced cAMP accumulation, which were fully prevented by montelukast and GF109203X, respectively. More importantly, GF109203X also prevented the attenuating effect of LTD₄ on isoproterenol-induced cAMP accumulation. In bronchial rings, both montelukast and GF109203X prevented the rightward displacement of the concentration-response curves to salbutamol induced by allergen challenge.

Conclusion

LTD₄ induces β₂-adrenoceptor desensitization in human airway smooth muscle cells, which is mediated through the activation of PKC. Allergen exposure of sensitized human bronchi may also cause a β₂-adrenoceptor desensitization through the involvement of the CysLT₁R-PKC pathway.