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Updated: Jul 23, 2026

In vitro Measurements of Tracheal Constriction Using Mice
Published on: June 25, 2012
TNF-alpha increases transcription of Galpha(i-2) in human airway smooth muscle cells
K Hotta1, C A Hirshman, C W Emala
1Department of Anesthesiology, College of Physicians and Surgeons of Columbia University, New York, New York 10032, USA.
Abstract:
Tumor necrosis factor-alpha (TNF-alpha) is a proinflammatory cytokine that has an important role in the regulation of airway smooth muscle tone and reactivity. We have shown previously that TNF-alpha upregulates the expression of Galpha(i-2) protein without significantly increasing G(s)alpha protein and enhances adenylyl cyclase inhibition by carbachol in cultured human airway smooth muscle cells (Hotta K, Emala CW, and Hirshman CA. Am J Physiol Lung Cell Mol Physiol 276: L405-L411, 1999). The present study was designed to investigate the molecular mechanisms by which TNF-alpha upregulates Galpha(i-2) protein in these cells. TNF-alpha pretreatment for 48 h increased the expression of Galpha(i-2) protein without significantly altering the Galpha(i-2) protein half-life (41.0 +/- 8.2 h for control and 46.8 +/- 5.2 h for TNF-alpha-treated cells). Inhibition of new protein synthesis by cycloheximide blocked the increase in Galpha(i-2) protein induced by TNF-alpha. Furthermore, TNF-alpha treatment for 12-24 h increased the steady-state level of Galpha(i-2) mRNA without significantly altering Galpha(i-2) mRNA half-life (9.0 +/- 0.75 h for control and 8.9 +/- 1.1 h for TNF-alpha-treated cells). The transcription inhibitor actinomycin D blocked the increase in Galpha(i-2) mRNA induced by TNF-alpha. These observations indicate that the increase in Galpha(i-2) protein induced by TNF-alpha is due to an increased rate of Galpha(i-2) protein synthesis, most likely as a consequence of the transcriptional increase in the steady-state levels of its mRNA.
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