Contribution of whole blood to the control of plasma asymmetrical dimethylarginine.
The endogenous nitric oxide (NO) synthase (NOS) inhibitor asymmetrical dimethylarginine (ADMA) is elevated in many patients and may contribute to the initiation and progression of their disease. While some mechanistic pathways have been identified, tissue-specific contributions to ADMA control remain unclear. We sought to determine if whole blood (WB) could participate in ADMA control ex vivo. Anesthetized male Sprague-Dawley rats underwent exsanguinations, and WB preparations were incubated at 37 degrees C for 5 h. ADMA and symmetrical dimethylarginine were analyzed by high-pressure liquid chromatography. Incubation of lysed red blood cell (RBC) supernatant yielded a significant decrease in ADMA that was blocked by 4124W, a synthetic inhibitor of dimethylarginine dimethylaminohydrolase, the only reported enzyme to hydrolyze ADMA. Hydrolysis of ADMA was diminished by addition of physiologically relevant concentrations of zinc (i.e., 20 microM). Conversely, when rat WB or WB supernatant was incubated at 37 degrees C, it liberated quantities of free ADMA (1-2 microM) that in vivo would likely have pathological consequences. Addition of arginine methyltransferase inhibitors to these incubations did not reduce ADMA release, indicating no dominant role for active protein methylation during these incubations. This ADMA liberation was significantly reduced by addition of protease inhibitors, indicating a dependence on peptide bond hydrolysis. Total ADMA (protein incorporated plus free) was determined by acid hydrolysis and found to be 43.18 +/- 4.79 microM in WB with approximately 95% of this in RBCs. These ex vivo data demonstrate the potential of blood to control the NO-NOS system by modulating free ADMA.
Billecke
Scott S
SS
7744 Medical Sciences Bldg. II, Dept. of Molecular and Integrative Physiology, Univ. of Michigan Medical School, Ann Arbor, MI 48109-0622, USA. lgdalecy@umich.edu
Kitzmiller
Laura A
LA
Northrup
Joseph J
JJ
Whitesall
Steven E
SE
Kimoto
Masumi
M
Hinz
Alia V
AV
D'Alecy
Louis G
LG
D000613:Aminobutyrates / Q000494:pharmacology; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000097:blood; D001769:Blood / Q000187:drug effects / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D006146:Guanidines / Q000494:pharmacology; D006973:Hypertension / Q000097:blood; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012435:S-Adenosylhomocysteine / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology
C514525:2-amino-4-(3-methylguanidino)butanoic acid; D000613:Aminobutyrates; D006146:Guanidines; C487735:dimethylarginine; D009569:Nitric Oxide; D001120:Arginine; D012435:S-Adenosylhomocysteine; D019001:Nitric Oxide Synthase; D015032:Zinc