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Am J Physiol Lung Cell Mol Physiol 280: L965-L973, 2001;
1040-0605/01 $5.00
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Vol. 280, Issue 5, L965-L973, May 2001

EETs relax airway smooth muscle via an EpDHF effect: BKCa channel activation and hyperpolarization

Catherine Benoit, Barbara Renaudondagger, Dany Salvail, and Eric Rousseau

Le Bilarium, Faculty of Medicine, Department of Physiology and Biophysics, University of Sherbrooke, Sherbrooke, Quebec, Canada J1H 5N4

Epoxyeicosatrienoic acids (EETs) are produced from arachidonic acid via the cytochrome P-450 epoxygenase pathway. EETs are able to modulate smooth muscle tone by increasing K+ conductance, hence generating hyperpolarization of the tissues. However, the molecular mechanisms by which EETs induce smooth muscle relaxation are not fully understood. In the present study, the effects of EETs on airway smooth muscle (ASM) were investigated using three electrophysiological techniques. 8,9-EET and 14,15-EET induced concentration-dependent relaxations of the ASM precontracted with a muscarinc agonist (carbamylcholine chloride), and these relaxations were partly inhibited by 10 nM iberiotoxin (IbTX), a specific large-conductance Ca2+-activated K+ (BKCa) channel blocker. Moreover, 3 µM 8,9- or 14,15-EET induced hyperpolarizations of -12 ± 3.5 and -16 ± 3 mV, with EC50 values of 0.13 and 0.14 µM, respectively, which were either reversed or blocked on addition of 10 nM IbTX. These results indicate that BKCa channels are involved in hyperpolarization and participate in the relaxation of ASM. In addition, complementary experiments demonstrated that 8,9- and 14,15-EET activate reconstituted BKCa channels at low free Ca2+ concentrations without affecting their unitary conductance. These increases in channel activity were IbTX sensitive and correlated well with the IbTX-sensitive hyperpolarization and relaxation of ASM. Together these results support the view that, in ASM, the EETs act through an epithelium-derived hyperpolarizing factorlike effect.

epoxyeicosatrienoic acids; bronchorelaxation; membrane potential; potassium conductance; eicosanoids; planar lipid bilayers; epithelium-derived hyperpolarizing factors; large-conductance calcium-activated potassium channel

dagger Deceased 30 October 2000.




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