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Am J Physiol Lung Cell Mol Physiol 275: L491-L501, 1998;
1040-0605/98 $5.00
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Vol. 275, Issue 3, L491-L501, September 1998

Prostanoids mediate IL-1beta -induced beta -adrenergic hyporesponsiveness in human airway smooth muscle cells

Johanne D. Laporte1, Paul E. Moore1, Reynold A. Panettieri2, Winfried Moeller3, Joachim Heyder3, and Stephanie A. Shore1

1 Physiology Program, Harvard School of Public Health, Boston, Massachusetts 02115; 2 Pulmonary and Critical Care Division, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19002-2209; and 3 GSF National Research Center for Environment and Health Institute of Inhalation Biology, Oberschleissheim, Germany

We have previously reported that pretreatment of cultured human airway smooth muscle (HASM) cells with interleukin-1beta (IL-1beta ) results in decreased beta -adrenergic responsiveness. The purpose of this study was to determine whether prostanoids released as a result of cyclooxygenase-2 (COX-2) induction by IL-1beta contribute to this effect of the cytokine. Confluent serum-deprived HASM cells were studied in passages 4-7. IL-1beta (20 ng/ml for 22 h) reduced the ability of the beta -agonist isoproterenol (Iso) to decrease stiffness of HASM cells as measured by magnetic twisting cytometry. The effect of IL-1beta on Iso-induced changes in cell stiffness was abolished by nonselective [indomethacin (Indo), 10-6 M] and selective (NS-398, 10-5 M) COX-2 inhibitors. Indo and NS-398 also inhibited both the increased basal cAMP and the decreases in Iso-stimulated cAMP production induced by IL-1beta . IL-1beta (20 ng/ml for 22 h) caused an increase in both basal (15-fold) and arachidonic acid (AA)-stimulated (10-fold) PGE2 release. Indo blocked basal and AA-stimulated PGE2 release in both control and IL-1beta -treated cells. NS-398 also markedly reduced basal and AA-stimulated PGE2 release in IL-1beta -treated cells but had no significant effect on AA-stimulated PGE2 release in control cells. Western blot analysis confirmed the induction of COX-2 by IL-1beta . Exogenously administered PGE2 (10-7 M, 22 h) caused a significant reduction in the ability of Iso to decrease cell stiffness, mimicking the effects of IL-1beta . Cycloheximide (10 µg/ml for 24 h), an inhibitor of protein synthesis, also abolished the effects of IL-1beta on Iso-induced cell stiffness changes and cAMP formation. In summary, our results indicate that IL-1beta significantly increases prostanoid release by HASM cells as a result of increased COX-2 expression. The prostanoids appear to contribute to beta -adrenergic hyporesponsiveness, perhaps by heterologous desensitization of the beta 2 receptor.

prostaglandin E2; cytoskeletal mechanics; indomethacin; NS-398; adenosine 3',5'-cyclic monophosphate; interleukin-1beta


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