title: Low concentrations of sphingosylphosphorylcholine enhance pulmonary artery vasoreactivity: the role of protein kinase C delta and Ca2+ entry.
abstract: Sphingosylphosphorylcholine (SPC) is a powerful vasoconstrictor, but in vitro its EC(50) is approximately 100-fold more than plasma concentrations. We examined whether subcontractile concentrations of SPC (<or=1 micromol/L) modulated vasoreactivity of rat intrapulmonary arteries using myography and measurement of intracellular [Ca(2+)]. SPC (1 micromol/L) had no effect on force or intracellular [Ca(2+)] on its own, but dramatically potentiated constrictions induced by approximately 25 mmol/L [K(+)], such that at 40 minutes, force and intracellular [Ca(2+)] (Fura PE3 340/380 ratio) were increased by 429+/-96% and 134+/-26%, respectively. The potentiation was stereospecific, apparent at concentrations >100 nmol/L of SPC, and independent of the endothelium, 2-aminoethoxydiphenylborane-sensitive Ca(2+) entry, and Rho kinase. It was abolished by the phospholipase C inhibitor U73122, the broad spectrum protein kinase C (PKC) inhibitor Ro31-8220, and the PKC delta inhibitor rottlerin, but not by Gö6976, which is ineffective against PKC delta. The potentiation could be attributed to enhancement of Ca(2+) entry. SPC also potentiated the responses to prostaglandin F(2 alpha) and U436619, which activate a 2-aminoethoxydiphenylborane sensitive nonselective cation channel in intrapulmonary arteries. In this case, potentiation was partially inhibited by diltiazem but abolished by 2-aminoethoxydiphenylborane, Ro31-8220, and rottlerin. SPC (1 micromol/L) caused translocation of PKC delta to the perinuclear region and cytoskeleton of cultured intrapulmonary artery smooth muscle cells. We present the novel finding that low, subcontractile concentrations of SPC potentiate Ca(2+) entry in intrapulmonary arteries through both voltage-dependent and independent pathways via a receptor-dependent mechanism involving PKC delta. This has implications for the physiological role of SPC, especially in cardiovascular disease, where SPC is reported to be elevated.
authors.affiliation: Department of Physiology, Henriette Raphael House, King's College London, Guy's Hospital Campus, London SE1 1UL, United Kingdom.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Snetkov
authors.forename: Gavin D
authors.initials: GD
authors.lastname: Thomas
authors.forename: Bonnie
authors.initials: B
authors.lastname: Teague
authors.forename: Richard M
authors.initials: RM
authors.lastname: Leach
authors.forename: Yasin
authors.initials: Y
authors.lastname: Shaifta
authors.forename: Greg A
authors.initials: GA
authors.lastname: Knock
authors.forename: Philip I
authors.initials: PI
authors.lastname: Aaronson
authors.forename: Jeremy P T
authors.initials: JP
authors.lastname: Ward
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002478:Cells, Cultured; D004110:Diltiazem / Q000494:pharmacology; D015237:Dinoprost / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004594:Electrophysiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007425:Intracellular Membranes / Q000378:metabolism; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000201:enzymology; D009994:Osmolar Concentration; D010767:Phosphorylcholine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D011651:Pulmonary Artery / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D013110:Sphingosine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000494:pharmacology; D014661:Vasoconstriction / Q000187:drug effects; D014666:Vasomotor System / Q000187:drug effects*
chemical_list: D002121:Calcium Channel Blockers; D004791:Enzyme Inhibitors; C005356:sphingosine phosphorylcholine; D010767:Phosphorylcholine; D015237:Dinoprost; D051745:Protein Kinase C-delta; D004110:Diltiazem; D013110:Sphingosine; D011188:Potassium; D002118:Calcium