Abstract
The effects of ouabain, an inhibitor of the plasmalemmal Na+/K+-ATPase activity, were examined in human isolated bronchus. Ouabain produced concentration-dependent contraction with −logEC50=7.16±0.11 and maximal effect of 67±4% of the response to acetylcholine (1 mM). Ouabain (10 μM)-induced contraction was epithelium-independent and was not depressed by inhibitors of cyclooxygenase and lipoxygenase, antagonists of muscarinic, histamine H1-receptors and α-adrenoceptors, or neuronal Na+ channel blockade. The inhibition of ouabain contraction in tissues bathed in K+-free medium, and the inhibition by ouabain of the K+-induced relaxation confirm that the contractile action of ouabain is mediated by inhibition of Na+/K+-ATPase. Furthermore, depolarization (16.4±0.9 mV) was observed in human isolated bronchus by intracellular microelectrode recording. Ouabain (10 μM)-induced contractions were abolished by a Ca2+-free solution but not by blockers of L-type Ca2+ channels. In human cultured bronchial smooth muscle cells, ouabain (10 μM) produced a sustained increase in [Ca2+]i (116±26 nM) abolished in Ca2+-free medium. Incubation with a Na+-free medium or amiloride (0.1 mM) markedly inhibited the spasmogenic effect of ouabain thus suggesting the role of Na+/Ca2+ exchange in ouabain contraction while selective inhibitors of Na+/H+-antiport, Na+/K+/Cl−-antiport, or protein kinase C had no effect. Ouabain (10 μM) failed to increase inositol phosphate accumulation in human bronchus. Ouabain (10 μM) did not alter bronchial responsiveness to acetylcholine or histamine but inhibited the relaxant effects of isoprenaline, forskolin, levcromakalim, or sodium nitroprusside. These results indicate that ouabain acts directly to produce contraction of human airway smooth muscle that depends on extracellular Ca2+ entry unrelated to L-type channels and involving the Na+/Ca2+-antiporter.
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Acknowledgements
The present work was supported in part by grants SAF2000–0144 and SAF2002–04667 from CICYT (Ministry of Science and Technology, Spain). The authors are indebted to the teams of the Services of Thoracic Surgery and Pathology of the Hospital La Fe and Hospital Clínico of the University of Valencia for making the human lung tissue available to us. We also thank V. Villagrasa, P. Santamaría and S. Martí for help with experiments.
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Cortijo, J., Sarria, B., Mata, M. et al. Effects of ouabain on human bronchial muscle in vitro. Naunyn-Schmiedeberg's Arch Pharmacol 368, 393–403 (2003). https://doi.org/10.1007/s00210-003-0818-0
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DOI: https://doi.org/10.1007/s00210-003-0818-0