Altered L-arginine metabolism results in increased nitric oxide release from uraemic endothelial cells.
Results regarding the nitric oxide (NO) system in uraemia are contradictory. L-arginine, the precursor of NO, is also metabolized by arginase to form ornithine and urea. In the present study, endothelial NO production and arginine metabolism in uraemia were assessed. In addition an in vivo model was used to examine excess consumption of NO in uraemia. NO and amino acid measurements were made from basal and stimulated (by bradykinin) uraemic and control endothelial cells. Reverse-transcriptase PCR was used to assess endothelial NO synthase (eNOS) and inducible NOS (iNOS) expression. Finally, aortae of uraemic rats were stained for nitrotyrosine (a marker of peroxynitrite). Basal uraemic cells produced more NO than the control cells. L-arginine levels were greater in uraemic (supernatants/cells), but ornithine levels were higher in control (supernatants/cells). Following stimulation, NO levels in supernatants were similar, but the rise in NO production was greater in control compared with uraemic cells; l-arginine levels still remained higher in uraemic supernatants/cells. Differences in ornithine concentration (supernatants/cells) disappeared following bradykinin stimulation, due to a rise in ornithine levels in the uraemic group. There was no difference in eNOS expression, nor was iNOS detected in either group. Only aortae from uraemic rats showed evidence for nitrotyrosine staining. These studies demonstrated increased basal NO release in uraemic endothelial cells, perhaps by inhibition of arginase and hence diversion of arginine to the NO pathway. The increased NO produced under basal conditions may be inactive due to excessive consumption, resulting in peroxynitrite formation. Interestingly, bradykinin appears to restore arginase activity in uraemia, resulting in normalization of NO production.
Thuraisingham
Raj C
RC
Anthony Raine Research Laboratory, St. Bartholomew's Hospital, Queen Mary and Westfield College, London EC1A 7BE, UK. r.c.thuraising@mds.qmw.ac.uk
Roberts
Norman B
NB
Wilkes
Mark
M
New
David I
DI
Mendes-Ribeiro
A Claudio
AC
Dodd
Susan M
SM
Yaqoob
Muhammad M
MM
D000328:Adult; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001011:Aorta / Q000378:metabolism; D001120:Arginine / Q000378:metabolism*; D001920:Bradykinin / Q000494:pharmacology; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009952:Ornithine / Q000378:metabolism; D051381:Rats; D011921:Rats, Inbred WKY; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014511:Uremia / Q000378:metabolism*
D000596:Amino Acids; D017077:Culture Media, Conditioned; D009569:Nitric Oxide; D001120:Arginine; D009952:Ornithine; D019001:Nitric Oxide Synthase; D001920:Bradykinin