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Published ahead of print on April 29, 2004, doi:10.1164/rccm.200308-1071OC

Am. J. Respir. Crit. Care Med., Volume 170, Number 2, July 2004, 154-161

A more recent version of this article appeared on July 15, 2004
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Submitted on August 1, 2003
Accepted on April 23, 2004

Differential Cytokine Gene Expression in the Diaphragm in Response to Strenuous Resistive Breathing

Theodoros Vassilakopoulos1*, Maziar Divangahi1, George Rallis1, Osama Kishta1, Basil Petrof1, Alain Comtois1, and Sabah N.A. Hussain1

1 Critical Care and Respiratory Divisions, Department of Medicine, McGill University Hospital Center, Montreal, Quebec, Canada; Meakins-Christie Laboratories, McGill University, Montreal, Quebec, Canada

* To whom correspondence should be addressed. E-mail: tvassilakopoulos{at}yahoo.com.

Strenuous resistive breathing induces plasma cytokines that do not originate from circulating monocytes. We hypothesized that cytokine production is induced inside the diaphragm in response to resistive loading. Anesthetized, tracheostomized, spontaneously breathing Sprague-Dawely rats were subjected to 1, 3 or 6 hours of inspiratory resistive loading, corresponding to 45-50% of the maximum inspiratory pressure. Unloaded sham operated rats breathing spontaneously served as controls. The diaphragm and the gastrocnemius muscles were excised at the end of the loading period and messenger ribonucleic acid expression of tumor necrosis factor-alpha and beta, interleukin-1alpha, interleukin -1{beta}, interleukin-2, interleukin -3, interleukin-4, interleukin -5, interleukin -6, interleukin-10, interferon-gamma and 2 housekeeping genes was analysed using multi-probe ribonuclease protection assay. Interleuhin-6, interleukin-1{beta} and to lesser extent tumour necrosis factor-alpha, interleukin-10, interferon-gamma and interleukin-4 were significantly increased in a time-dependent fashion in the diaphragms but not the gastrocnemius of loaded animals or in the diaphragm of control animals. Elevation of protein levels of interleukin-6 and interleukin-1{beta} in the diaphragm of loaded animals was confirmed with immunoblotting. Immunostaining revealed interleukin-6 protein localization inside diaphragmatic muscle fibers. We conclude that increased ventilatory muscle activity during resistive loading induces differential elevation of pro- and anti-inflammatory cytokine gene expression in the ventilatory muscles.
Key words: respiratory muscles, interleukin, loaded breathing, ribonuclease protection assay




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