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1 Department of Pharmacology, University of Sydney, Sydney, NSW, Australia; Department of Internal Medicine and Research, University Hospital Basel, Basel, Switzerland
2 Department of Pharmacology, University of Sydney, Sydney, NSW, Australia
* To whom correspondence should be addressed. E-mail: Stephen.Carlin{at}unibas.ch.
We investigated the chemotactic action of PDGF and urokinase on human airway smooth muscle (HASM) cells in culture. Cells were put in collagen-coated transwells with 8 µm perforations, incubated for four hours with test compounds, then fixed, stained, and counted as migrated nuclei by microscopy. Cells from all culture conditions showed some basal migration (migration in the absence of stimuli during the assay), but cells pre-incubated for 24 h in 10% FBS or 20 ng/mL PDGF showed higher basal migration than cells quiesced in 1 % FBS. PDGFBB, PDGFAA and PDGFAB were all chemotactic when added during the assay. PDGF chemotaxis was blocked by; the PI3-kinase inhibitor LY294002, the MEK inhibitor U0126, PGE2, formoterol, pertussis toxin, and the Rho kinase inhibitor Y27632. Urokinase alone had no stimulatory effect on migration of quiescent cells, but caused a dose-dependent potentiation of chemotaxis toward PDGF. Urokinase also potentiated the elevated basal migration of cells pre-treated in 10% FBS or PDGF. This potentiating effect of urokinase appears to be novel. We conclude that PDGF and similar cytokines may be important factors in airway remodelling by redistribution of smooth muscle cells during inflammation, and that urokinase may be important in potentiating the response.
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