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B Mediate COX-2 Expression in Human Airway Myocytes
1 Department of Pharmacology, University of Nevada School of Medicine, Reno, NV, USA
2 Department of Cell and Molecular Biology Program, University of Nevada School of Medicine, Reno, NV, USA
3 Department of Pharmacology, University of Nevada School of Medicine, Reno, NV, USA; Department of Cell and Molecular Biology Program, University of Nevada School of Medicine, Reno, NV, USA
* To whom correspondence should be addressed. E-mail: cas{at}med.unr.edu.
We have previously demonstrated that p38 and extracellular signal-regulated protein kinase (ERK) mitogen-activated protein kinases (MAPK) are components of pro-inflammatory induced cytokine expression in human airway myocytes. The experiments described here further these studies by examining p38 MAPK and NF-
B regulation of cyclooxygenase-2 (COX-2) expression in response to a complex inflammatory stimulus consisting of 10 ng/ml interleukin (IL)-1
, tumor necrosis factor (TNF)
and interferon (IFN)
. COX-2 expression was induced with this stimulus in a time-dependent manner, with maximal expression seen 12-20 hours after treatment. Semi-quantitative RT-PCR and immunoblotting experiments demonstrate decreased COX-2 expression following treatment with the p38 MAPK inhibitor SB203580 (25 µM), or the proteosome inhibitor MG-132 (1 µM). SB20380 did not affect cytokine-stimulated IB degradation, NF-B nuclear binding activity or NF-B dependent signaling from the COX-2 promoter, indicating that p38 MAPK and NF-B affect COX-2 expression via separate signaling pathways. SB203580, but not MG-132, also increased the initial rate of COX-2 mRNA decay, indicating p38 MAPK, but not NF-B, participates in the regulation of COX-2 mRNA stability. These findings suggest that while p38 MAPK and NF-B signaling regulate steady-state levels of COX-2 expression, p38 MAPK additionally affects stability of COX-2 mRNA in cytokine-stimulated human airway myocytes.
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