Tachykinin-1 receptor stimulates proinflammatory gene expression in lung epithelial cells through activation of NF-kappaB via a G(q)-dependent pathway.
The respiratory tract is innervated by irritant-responsive sensory nerves, which, on stimulation, release tachykinin neuropeptides in the lung. Tachykinins modulate inflammatory responses to injury by binding to tachykinin (neurokinin) receptors present on various pulmonary cell types. In the present study, the activation of the proinflammatory transcription factor NF-kappaB in lung epithelial cells was investigated as a mechanism by which tachykinins stimulate inflammatory processes. In A549 human lung epithelial cells transfected with the tachykinin-1 receptor (Tacr1), treatment with the Tacr1 ligand substance P (SP) resulted in NF-kappaB activation, as judged by transcription of an NF-kappaB-luciferase reporter gene and production of interleukin-8, a chemokine whose expression is upregulated by NF-kappaB. SP caused a dose-dependent activation of NF-kappaB that was inhibited by the selective Tacr1 antagonist RP67580. Tacr1 is a G protein-coupled receptor capable of activating both the G(q) and G(s) families of G proteins. Expression of inhibitory peptides and constitutively active G protein mutants revealed that G(q) signaling was both necessary for Tacr1-induced NF-kappaB activation and sufficient for NF-kappaB activation in the absence of any other treatment. Treatment with pharmacological inhibitors to investigate events downstream of G(q) revealed that Tacr1-induced NF-kappaB activation proceeded through an intracellular signaling pathway that was dependent on phospholipase C, calcium, Ras, Raf-1, MEK, Erk, and proteasome function. These results identify intracellular signaling mechanisms that underlie the proinflammatory effects of tachykinins, which previously have been implicated in lung injury and disease.
Williams
Ronald
R
Section of Pulmonology, Department of Pediatrics, Tulane University Health Sciences Center, New Orleans, Louisiana 70112, USA.
Zou
Xiaoyan
X
Hoyle
Gary W
GW
D000818:Animals; D004789:Enzyme Activation / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism*; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D007976:Leupeptins / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D008168:Lung / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D016328:NF-kappa B / Q000235:genetics*; D061988:Proteasome Inhibitors; D018040:Receptors, Neurokinin-1 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013373:Substance P / Q000494:pharmacology; D014162:Transfection
D018836:Inflammation Mediators; D007976:Leupeptins; D016328:NF-kappa B; D061988:Proteasome Inhibitors; D018040:Receptors, Neurokinin-1; D013373:Substance P; D008156:Luciferases; D048049:Extracellular Signal-Regulated MAP Kinases; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D019205:GTP-Binding Protein alpha Subunits, Gs; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde