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Am J Physiol Lung Cell Mol Physiol 277: L606-L615, 1999;
1040-0605/99 $5.00
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Vol. 277, Issue 3, L606-L615, September 1999

LPS-induced depolymerization of cytoskeleton and its role in TNF-alpha production by rat pneumocytes

Noritaka Isowa1, Alexandre M. Xavier1, Ewa Dziak2, Michal Opas2, Donna I. McRitchie3, Arthur S. Slutsky4, Shaf H. Keshavjee1,3, and Mingyao Liu1,3

1 Thoracic Surgery Research Laboratory, Toronto General Hospital, University Health Network, and Departments of 2 Anatomy and Cell Biology, 3 Surgery, and 4 Medicine, University of Toronto, Toronto, Ontario, Canada M5G 2C4

Lipopolysaccharide (LPS) polymerizes microfilaments and microtubules in macrophages and monocytes. Disrupting microfilaments or microtubules with cytochalasin D (CytoD) or colchicine can suppress LPS-induced tumor necrosis factor-alpha (TNF-alpha ) gene expression and protein production from these cells. We have recently demonstrated that primary cultured rat alveolar epithelial cells can produce TNF-alpha on LPS stimulation. In the present study, we found that the LPS-induced increase in TNF-alpha mRNA level and protein production in alveolar epithelial cells was not inhibited by CytoD or colchicine (1 nM to 10 µM). In fact, LPS-induced TNF-alpha production was further enhanced by CytoD (1-10 µM) and inhibited by jasplakinolide, a polymerizing agent for microfilaments. Immunofluorescent staining and confocal microscopy showed that LPS (10 µg/ml) depolymerized microfilaments and microtubules within 15 min, which was prolonged until 24 h for microfilaments. These results suggest that the effects of LPS on the cytoskeleton and the role of the cytoskeleton in mediating TNF-alpha production in alveolar epithelial cells are opposite to those in immune cells. This disparity may reflect the different roles between nonimmune and immune cells in host defense.

tumor necrosis factor-alpha ; cytokines; lipopolysaccharide; microfilament; microtubule


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