title: Radiochemical high-performance liquid chromatography detection of arginine metabolism in human endothelial cells.
abstract: Arginine is a semi-essential amino acid that plays an important role in the regulation of metabolic processes associated with several pathological/physiological conditions. In the vasculature, it mainly exerts its biological functions as a substrate of two alternative pathways: the conversion to nitric oxide (NO) by nitric oxide synthase (NOS) and the breakdown to urea and ornithine by arginase. To determine arginine metabolism, in the current study we propose an original radiochemical technique that allows the simultaneous monitoring of NOS and arginase activation within intact cells. Taking advantage of this method, we show here the consequences of different experimental conditions known to modulate endothelial homeostasis on arginine metabolism.
authors.affiliation: Dipartimento di Medicina Sperimentale, Sezione di Patologia Generale e Clinica, Università degli Studi di Parma, 43125 Parma, Italy.
authors.forename: Amelia
authors.initials: A
authors.lastname: Barilli
authors.forename: Rossana
authors.initials: R
authors.lastname: Visigalli
authors.forename: Bianca Maria
authors.initials: BM
authors.lastname: Rotoli
authors.forename: Ovidio
authors.initials: O
authors.lastname: Bussolati
authors.forename: Gian Carlo
authors.initials: GC
authors.lastname: Gazzola
authors.forename: Alessandro
authors.initials: A
authors.lastname: Parolari
authors.forename: Valeria
authors.initials: V
authors.lastname: Dall'Asta
mesh_terms: D001119:Arginase / Q000378:metabolism*; D001120:Arginine / Q000378:metabolism*; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D009952:Ornithine / Q000378:metabolism; D011854:Radiochemistry / Q000379:methods; D020123:Sirolimus / Q000494:pharmacology; D014316:Tritium; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014508:Urea / Q000378:metabolism
chemical_list: D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D014316:Tritium; D009569:Nitric Oxide; D014508:Urea; D001120:Arginine; D009952:Ornithine; D052250:Nitric Oxide Synthase Type III; D001119:Arginase; D020123:Sirolimus