Enhanced XOR activity in eNOS-deficient mice: Effects on the nitrate-nitrite-NO pathway and ROS homeostasis.
Xanthine oxidoreductase (XOR) is generally known as the final enzyme in purine metabolism and as a source of reactive oxygen species (ROS). In addition, this enzyme has been suggested to mediate nitric oxide (NO) formation via reduction of inorganic nitrate and nitrite. This NO synthase (NOS)-independent pathway for NO generation is of particular importance during certain conditions when NO bioavailability is diminished due to reduced activity of endothelial NOS (eNOS) or increased oxidative stress, including aging and cardiovascular disease. The exact interplay between NOS- and XOR-derived NO generation is not fully elucidated yet. The aim of the present study was to investigate if eNOS deficiency is associated with changes in XOR expression and activity and the possible impact on nitrite, NO and ROS homeostasis. Plasma levels of nitrate and nitrite were similar between eNOS deficient (eNOS-/-) and wildtype (wt) mice. XOR activity was upregulated in eNOS-/-compared with wt, but not in nNOS-/-, iNOS-/-or wt mice treated with the non-selective NOS inhibitor L-NAME. Following an acute dose of nitrate, plasma nitrite increased more in eNOS-/-compared with wt, and this augmented response was abolished by the selective XOR inhibitor febuxostat. Livers from eNOS-/-displayed higher nitrite reducing capacity compared with wt, and this effect was attenuated by febuxostat. Dietary supplementation with nitrate increased XOR expression and activity, but concomitantly reduced superoxide generation. The latter effect was also seen in vitro after nitrite administration. Treatment with febuxostat elevated blood pressure in eNOS-/-, but not in wt mice. A high dose of dietary nitrate reduced blood pressure in naïve eNOS-/-mice, and again this effect was abolished by febuxostat. In conclusion, eNOS deficiency is associated with an upregulation of XOR facilitating the nitrate-nitrite-NO pathway and decreasing the generation of ROS. This interplay between XOR and eNOS is proposed to play a significant role in NO homeostasis and blood pressure regulation.
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Maria
M
Peleli
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Christa
C
Zollbrecht
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Marcelo F
MF
Montenegro
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Michael
M
Hezel
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Jianghong
J
Zhong
Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden.
Erik G
EG
Persson
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Rikard
R
Holmdahl
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Eddie
E
Weitzberg
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden. Electronic address: Jon.Lundberg@ki.se.
Jon O
JO
Lundberg
Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden. Electronic address: Mattias.Carlstrom@ki.se.
Mattias
M
Carlström
D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000069465:Febuxostat / Q000494:pharmacology; D005786:Gene Expression Regulation; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009566:Nitrates / Q000097:blood / Q000494:pharmacology; D009569:Nitric Oxide / Q000097:blood*; D052248:Nitric Oxide Synthase Type I / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics*; D052250:Nitric Oxide Synthase Type III / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics*; D009573:Nitrites / Q000097:blood / Q000494:pharmacology; D010084:Oxidation-Reduction; D015398:Signal Transduction; D013481:Superoxides / Q000378:metabolism; D014968:Xanthine Dehydrogenase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
D004791:Enzyme Inhibitors; D009566:Nitrates; D009573:Nitrites; D000069465:Febuxostat; D013481:Superoxides; D009569:Nitric Oxide; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496321:Nos1 protein, mouse; C496317:Nos2 protein, mouse; C496315:Nos3 protein, mouse; D014968:Xanthine Dehydrogenase; D019331:NG-Nitroarginine Methyl Ester
Endothelial nitric oxide synthase; Febuxostat; Hypertension; Nitrate; Nitric oxide; Nitrite; Oxidative stress; Reactive oxygen species; Superoxide; Uric acid; Xanthine oxidase; Xanthine oxidoreductase