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responses in cultured airway
smooth muscle cells
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)-1
causes -adrenergic hyporesponsiveness in cultured human
airway smooth muscle (HASM) cells by increasing cyclooxygenase (COX)-2
expression. The purpose of this study was to determine whether p38
mitogen-activated protein (MAP) kinase is involved in these events.
IL-1 (2 ng/ml for 15 min) increased p38 phosphorylation fourfold.
The p38 inhibitor SB-203580 (3 µM) decreased IL-1-induced COX-2 by
70 ± 7% (P < 0.01). SB-203580 had no effect on
PGE2 release in control cells but caused a significant
(70-80%) reduction in PGE2 release in IL-1-treated
cells. IL-1 increased the binding of nuclear proteins to the
oligonucleotides encoding the consensus sequences for activator protein
(AP)-1 and nuclear factor (NF)-
B, but SB-203580 did not affect this
binding, suggesting that the mechanism of action of p38 was not through
AP-1 or NF-B activation. The NF-B inhibitor MG-132 did not alter
IL-1-induced COX-2 expression, indicating that NF-B activation is
not required for IL-1-induced COX-2 expression in HASM cells.
IL-1 attenuated isoproterenol-induced decreases in HASM stiffness as
measured by magnetic twisting cytometry, and SB-203580 abolished this
effect. These results are consistent with the hypothesis that p38 is
involved in the signal transduction pathway through which IL-1
induces COX-2 expression, PGE2 release, and -adrenergic hyporesponsiveness.
mitogen-activated protein; interleukin-1; human airway smooth
muscle; SB-203580; nuclear factor-B; activator protein-1; prostaglandin E2; cyclooxygenase-2; -adrenergic
responsiveness; cytoskeletal mechanics; magnetic twisting cytometry
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