title: Formation of endothelium-derived relaxing factor in porcine kidney epithelial LLC-PK1 cells: an intra- and intercellular messenger for activation of soluble guanylate cyclase.
abstract: Oxytocin, bradykinin, melittin and A23187 increased cyclic GMP levels through activation of soluble guanylate cyclase in cultured porcine kidney epithelial cells, LLC-PK1. NG-monomethyl-L-arginine, an inhibitor of endothelium-derived relaxing factor/nitric oxide formation, decreased both basal and stimulated levels of cyclic GMP in a concentration-dependent manner. L-Arginine, but not D-arginine, augmented basal as well as stimulated levels of cyclic GMP and prevented the inhibition induced by NG-monomethyl-L-arginine. Similar effects of L-arginine were also observed with L-argininamide, L-arginine ethyl ester, L-arginine methyl ester and the dipeptide L-arginyl-L-aspartic acid. NG-monomethyl-L-arginine did not affect cyclic GMP accumulation induced by sodium nitroprusside, an activator of soluble guanylate cyclase, and atrial natriuretic factor, an activator of particulate guanylate cyclase. Stimulatory effects of oxytocin, glyceryl trinitrate, sodium nitroprusside, bradykinin, melittin and A23187 on cyclic GMP accumulation were enhanced with superoxide dismutase and diminished with oxyhemoglobin. However, atrial natriuretic factor-induced cyclic GMP accumulation was not affected. Furthermore, endothelium derived relaxing factor-like activity was detected in the conditioned medium from LLC-PK1 cells stimulated with oxytocin. Based on these data, we conclude that endothelium-derived relaxing factor is produced in this cell type and participates in the regulatory mechanism of cyclic GMP formation as an intra- and intercellular messenger for activation of soluble guanylate cyclase.
authors.affiliation: Department of Pharmacology, Northwestern University Medical School, Chicago, Illinois.
authors.forename: K
authors.initials: K
authors.lastname: Ishii
authors.forename: B
authors.initials: B
authors.lastname: Chang
authors.forename: J F
authors.initials: JF
authors.lastname: Kerwin
authors.forename: F L
authors.initials: FL
authors.lastname: Wagenaar
authors.forename: Z J
authors.initials: ZJ
authors.lastname: Huang
authors.forename: F
authors.initials: F
authors.lastname: Murad
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media; D006152:Cyclic GMP / Q000378:metabolism; D004789:Enzyme Activation; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005333:Fetus / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006162:Guanylate Cyclase / Q000378:metabolism*; D007365:Intercellular Junctions / Q000378:metabolism; D007424:Intracellular Fluid / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D008168:Lung; D009569:Nitric Oxide / Q000096:biosynthesis*; D010121:Oxytocin / Q000494:pharmacology; D013552:Swine
chemical_list: D003470:Culture Media; D009569:Nitric Oxide; D010121:Oxytocin; D006162:Guanylate Cyclase; D006152:Cyclic GMP