title: Pharmacological characterization of mucin secretion from CHO-K1-hNK(1)R cells.
abstract: Numerous respiratory diseases increase mucin secretion from human airways. Several investigators hypothesize that mucin secretion from airway epithelium is NK(1)-receptor mediated. We have developed a mucin secretion assay using CHO-K1 cells transfected with the human NK(1)receptor (CHO-K1-hNK(1)R) that respond to NK(1)-specific agonists. Cells were labeled with [(3)H]-glucosamine and stimulated with agonists including Ac-[Arg(6), Sar(9), Met(O(2))(11)] Substance P(6-11) (ASMSP; NK(1)-specific), [beta-Ala(8)]-Neurokinin A(4-10) (BANK; NK(2)-specific), or human neutrophil elastase (HNE). Basal mucin secretion from CHO-K1-hNK(1)R and non-transfected cells was similar. Stimulation of CHO-K1-hNK(1)R, but not CHO-K1, with ASMSP or BANK concentration-dependently increased mucin secretion (pD(2)value[Emax] = 8.9(1)+/-0.1(3)[175%] and 7.56+/-0.05[100%], respectively). SR140333 (NK(1)antagonist), but not SR48968 (NK(2)antagonist), decreased ASMSP- and BANK-induced mucin release from CHO-K1-hNK(1)R. In these cells, endothelin-1, angiotensin II, serotonin, phenylephrine, senktide, and methacholine showed negligible effects on mucin secretion. A similar lack of effect of these agonists was observed in non-transfected CHO-K1 cells. HNE increased mucin release four to five fold in both cell types. These studies demonstrate that stimulation of CHO- K1-hNK(1)R with ASMSP and BANK causes robust and NK(1)-selective mucin release.
authors.affiliation: Respiratory Inflammatory, and Neurological Diseases Research Dept, Wilmington, DE, USA.
authors.forename: R
authors.initials: R
authors.lastname: Caccese
authors.forename: A
authors.initials: A
authors.lastname: Graham
authors.forename: R
authors.initials: R
authors.lastname: Bialecki
mesh_terms: D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D016466:CHO Cells; D006224:Cricetinae; D019332:Endothelin-1 / Q000494:pharmacology; D005944:Glucosamine / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D016210:Methacholine Chloride / Q000494:pharmacology; D009077:Mucins / Q000378:metabolism*; D018721:Muscarinic Agonists / Q000494:pharmacology; D015288:Neurokinin A / Q000031:analogs & derivatives / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D010656:Phenylephrine / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D011812:Quinuclidines / Q000494:pharmacology; D018040:Receptors, Neurokinin-1 / Q000235:genetics* / Q000378:metabolism*; D012701:Serotonin / Q000494:pharmacology; D013373:Substance P / Q000031:analogs & derivatives / Q000494:pharmacology; D014162:Transfection; D014316:Tritium; D014662:Vasoconstrictor Agents / Q000494:pharmacology
chemical_list: D001549:Benzamides; D019332:Endothelin-1; D009077:Mucins; D018721:Muscarinic Agonists; D010446:Peptide Fragments; D010880:Piperidines; D011812:Quinuclidines; D018040:Receptors, Neurokinin-1; D014662:Vasoconstrictor Agents; C082580:substance P(4-11), beta Ala(4)-Ser(9)-Met(O2)(11)-; D016210:Methacholine Chloride; D014316:Tritium; D000804:Angiotensin II; C064218:neurokinin A (4-10), beta-Ala(8)-; C085215:SR 140333; D010656:Phenylephrine; D012701:Serotonin; D013373:Substance P; C073839:SR 48968; D015288:Neurokinin A; D005944:Glucosamine