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J. Biol. Chem., Vol. 278, Issue 21, 18868-18876, May 23, 2003

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CCAAT/Enhancer-binding Protein and Activator Protein-1 Transcription Factors Regulate the Expression of Interleukin-8 through the Mitogen-activated Protein Kinase Pathways in Response to Mechanical Stretch of Human Airway Smooth Muscle Cells^*

Ashok Kumar, Alan J. Knox  and Aladin M. Boriek 

From the Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, Division of Respiratory Medicine, City Hospital, University of Nottingham, Nottingham NG7 2RD, United Kingdom

Here we investigated the mechanisms by which mechanical stretch regulates the production of IL-8 in primary human airway smooth muscle cells (HASMC). Bronchial HASMC were subjected to cyclic mechanical stretch (12%, 1 Hz) using the computer-controlled Flexcell Strain system. Mechanical stretch increased IL-8 mRNA expression and protein production. Cyclic stretch of HASMC also increased the kinase activities of ERK1/2, JNK1, p38, and the DNA binding activities of AP-1 and C/EBP transcription factors with little effect on NF-B. The inhibition of AP-1 and C/EBP transcriptional activities blocked the production of IL-8 in culture supernatants. Furthermore, the inhibition of ERK1/2 and p38 but not JNK1 caused a significant down-regulation in the expression and production of IL-8 in response to cyclic stretch. Although protein tyrosine kinases were required for the activation of both ERK1/2 and p38 kinase, stretch-activated channels, small GTPase proteins, and extracellular Ca²⁺ influx were required only for the activation of p38 kinase whereas phosphoinositide 3-kinase was needed for ERK1/2 activation. In addition, the phosphorylation of ERK1/2 was essential for the activation of AP-1 whereas p38 MAP kinase was needed for the activation of C/EBP. Our data demonstrate that the cyclic stretch of HASMC causes the increased production of IL-8 by activating the AP-1 and C/EBP transcription factors through the activation of ERK1/2 and p38 kinase signaling pathways.

Received for publication, December 16, 2002 , and in revised form, January 24, 2003.

To whom correspondence should be addressed: Pulmonary and Critical Care, Suite 520B, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Fax: 713-798-3619; E-mail: boriek{at}bcm.tmc.edu.

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