Levcromakalim- and Isoprenaline-induced Relaxation of Human Isolated Airways—Role of the Epithelium and of K+ Channel Activation

Summary: In this study we have investigated the mechanism of action of levcromakalim and isoprenaline in human isolated airways with respect to the K+ channels they activate and the possibility that these smooth muscle relaxants activate K+ channels on the airway epithelium. Mechanical removal of the epithelial layer (mean percentage of epithelium present 20±3%, n=20 tissues) did not affect the relaxation responses to levcromakalim or isoprenaline, either in terms of maximal relaxation or sensitivity. Whilst having no effect on isoprenaline-induced relaxation, studied from basal tone, the ATP-sensitive K+ channel blocker BRL 31660 (10, 30 and 50 μM) reduced relaxation responses induced (from basal tone) by levcromakalim from 74±6% (of the maximal response to isoprenaline) to 48±12% (n=7), 9±9% (n=4) and 0 (n=4), respectively. Charybdotoxin, a blocker of high conductance Ca2+-activated K+ channels, at concentrations of 30 and 100 nM, had no effect on either levcromakalim- or isoprenaline-induced relaxation responses and yet charybdotoxin was active at KCa channels in outside-out patches of hippocampal granule cells. Moreover, tetraethylammonium (10 mM) inhibited neither isoprenaline- nor levcromakalim-induced relaxation. This study has demonstrated that the relaxation responses elicited in human bronchus to isoprenaline and levcromakalim are likely to be the result of direct effects on the smooth muscle with no contribution from epithelial receptors or K+ channels. The actions of levcromakalim appear to be mediated only via activation of KATP channels. Further, we have made the important observation that, under the experimental conditions of our study, isoprenaline does not activate the KCa channel to produce relaxation in human bronchus.