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Am J Physiol Lung Cell Mol Physiol 277: L943-L951, 1999;
1040-0605/99 $5.00
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Vol. 277, Issue 5, L943-L951, November 1999

Role of ERK MAP kinases in responses of cultured human airway smooth muscle cells to IL-1beta

Johanne D. Laporte1, Paul E. Moore1, Joseph H. Abraham1, Geoffrey N. Maksym1, Ben Fabry1, Reynold A. Panettieri Jr.2, and Stephanie A. Shore1

1 Physiology Program, Harvard School of Public Health, Boston, Massachusetts 02115; and 2 Pulmonary and Critical Care Division, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104

We have previously reported that interleukin (IL)-1beta causes beta -adrenergic hyporesponsiveness in cultured human airway smooth muscle cells by increasing cyclooxygenase-2 (COX-2) expression and prostanoid formation. The purpose of this study was to determine whether extracellular signal-regulated kinases (ERKs) are involved in these events. Levels of phosphorylated ERK (p42 and p44) increased 8.3- and 13-fold, respectively, 15 min after treatment with IL-1beta (20 ng/ml) alone. Pretreating cells with the mitogen-activated protein kinase kinase inhibitor PD-98059 or U-126 (2 h before IL-1beta treatment) decreased ERK phosphorylation. IL-1beta (20 ng/ml for 22 h) alone caused a marked induction of COX-2 and increased basal PGE2 release 28-fold (P < 0.001). PD-98059 (100 µM) and U-126 (10 µM) each decreased COX-2 expression when administered before IL-1beta treatment. In control cells, PD-98059 and U-126 had no effect on basal or arachidonic acid (AA; 10 µM)-stimulated PGE2 release, but both inhibitors caused a significant decrease in bradykinin (BK; 1 µM)-stimulated PGE2 release, consistent with a role for ERK in the activation of phospholipase A2 by BK. In IL-1beta -treated cells, prior administration of PD-98059 caused 81, 92 and 40% decreases in basal and BK- and AA-stimulated PGE2 release, respectively (P < 0.01), whereas administration of PD-98059 20 h after IL-1beta resulted in only 38 and 43% decreases in basal and BK-stimulated PGE2 release, respectively (P < 0.02) and had no effect on AA-stimulated PGE2 release. IL-1beta attenuated isoproterenol-induced decreases in human airway smooth muscle stiffness as measured by magnetic twisting cytometry, and PD-98059 or U-126 abolished this effect in a concentration-dependent manner. These results are consistent with the hypothesis that ERKs are involved early in the signal transduction pathway through which IL-1beta induces PGE2 synthesis and beta -adrenergic hyporesponsiveness and that ERKs act by inducing COX-2 and activating phospholipase A2.

extracellular signal-regulated kinase; mitogen-activated protein; interleukin-1beta ; prostaglandin E2; beta -adrenergic responses; PD-98059; U-126; magnetic twisting cytometry; cyclooxygenase


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