Impaired nitric oxide production in coronary endothelial cells of the spontaneously diabetic BB rat is due to tetrahydrobiopterin deficiency.
Endothelial cells (EC) from diabetic BioBreeding (BB) rats have an impaired ability to produce NO. This deficiency is not due to a defect in the constitutive isoform of NO synthase in EC (ecNOS) or alterations in intracellular calcium, calmodulin, NADPH or arginine levels. Instead, ecNOS cannot produce sufficient NO because of a deficiency in tetrahydrobiopterin (BH(4)), a cofactor necessary for enzyme activity. EC from diabetic rats exhibited only 12% of the BH(4) levels found in EC from normal animals or diabetes-prone animals which did not develop disease. As a result, NO synthesis by EC of diabetic rats was only 18% of that for normal animals. Increasing BH(4) levels with sepiapterin increased NO production, suggesting that BH(4) deficiency is a metabolic basis for impaired endothelial NO synthesis in diabetic BB rats. This deficiency is due to decreased activity of GTP-cyclohydrolase I, the first and rate-limiting enzyme in the de novo biosynthesis of BH(4). GTP-cyclohydrolase activity was low because of a decreased expression of the protein in the diabetic cells.
Meininger
C J
CJ
Cardiovascular Research Institute and Department of Medical Physiology, The Texas A&M University System Health Science Center, College Station, TX 77843-1114, USA. cjm@tamu.edu
Marinos
R S
RS
Hatakeyama
K
K
Martinez-Zaguilan
R
R
Rojas
J D
JD
Kelly
K A
KA
Wu
G
G
D000818:Animals; D001120:Arginine / Q000737:chemistry; D001708:Biopterins / Q000031:analogs & derivatives* / Q000172:deficiency* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003920:Diabetes Mellitus / Q000378:metabolism* / Q000473:pathology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D006136:GTP Cyclohydrolase / Q000378:metabolism; D015151:Immunoblotting; D007700:Kinetics; D009249:NADP / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D020033:Protein Isoforms; D011621:Pteridines / Q000494:pharmacology; D011622:Pterins; D051381:Rats; D011922:Rats, Mutant Strains / Q000378:metabolism*
D002147:Calmodulin; D020033:Protein Isoforms; D011621:Pteridines; D011622:Pterins; D001708:Biopterins; D009569:Nitric Oxide; D009249:NADP; D001120:Arginine; C016727:sepiapterin; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D006136:GTP Cyclohydrolase; C003402:sapropterin; D002118:Calcium