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Mechanisms Underlying Erythrocyte and Endothelial Nitrite Reduction to Nitric Oxide in Hypoxia: Role for Xanthine Oxidoreductase and Endothelial Nitric Oxide Synthase
Open accessAbstract
Reduction of nitrite (NO 2−) provides a major source of nitric oxide (NO) in the circulation, especially in hypoxemic conditions. Our previous studies suggest that xanthine oxidoreductase (XOR) is an important nitrite reductase in the heart and kidney. Herein, we have demonstrated that conversion of nitrite to NO by blood vessels and RBCs was enhanced in the presence of the XOR substrate xanthine (10 μmol/L) and attenuated by the XOR inhibitor allopurinol (100 μmol/L) in acidic and hypoxic conditions only. Whereas endothelial nitric oxide synthase (eNOS) inhibition had no effect on vascular nitrite reductase activity, in RBCs L-NAME, L-NMMA, and l-arginine inhibited nitrite-derived NO production by >50% (P<0.01) at pH 7.4 and 6.8 under hypoxic conditions. Western blot and immunohistochemical analysis of RBC membranes confirmed the presence of eNOS and abundant XOR on whole RBCs. Thus, XOR and eNOS are ideally situated on the membranes of RBCs and blood vessels to generate intravascular vasodilator NO from nitrite during ischemic episodes. In addition to the proposed role of deoxyhemoglobin, our findings suggest that the nitrite reductase activity within the circulation, under hypoxic conditions (at physiological pH), is mediated by eNOS; however, as acidosis develops, a substantial role for XOR becomes evident.
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“Other medications, including xanthine oxidase (XOR) inhibitors (e.g., allopurinol) and drugs that affect salivary flow or gastrointestinal motility, may further modify nitrate bioactivation [17,272].”
From Dietary Nitrate-Rich Vegetables as Natural Modulators of Health: Mechanisms and Benefits in Ageing Populations (2026) · cites this paper as [272]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Furthermore, the process was demonstrated to be dependent on xanthine oxidoreductase (XOR)”
From Vascular effects of dietary nitrate (as found in green leafy vegetables and beetroot) via the nitrate-nitrite-nitric oxide pathway (2013) · cites this paper as [125]
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In the citing paper
“eNOS can also bioactivate nitrite under anoxic and/or acidic conditions.102,103”
From Nitrate-Nitrite-Nitric Oxide Pathway: Implications for Anesthesiology and Intensive Care (2010) · cites this paper as [102]
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In the citing paper
“A similar pathway exists in the pulmonary vasculature in which mitochondria-generated oxidants signal the conserved phenomenon of hypoxic vasoconstriction (995).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [995]
Experimental — accuracy not yet measuredIn the citing paper
“Other proposed nitrite reductases catalyzing NO formation from nitrite under hypoxic conditions are carbonic anhydrase (1), xanthine oxidase (68), or eNOS (189), which are all present in RBCs.”
From Red Blood Cell Function and Dysfunction: Redox Regulation, Nitric Oxide Metabolism, Anemia (2017) · cites this paper as [189]
In the citing paper
“The NO-like effects of dietary nitrate range from lowering of blood pressure 14,38 to a reduction in experimental myocardial infarct size, 33 inhibition of platelet aggregation, 34 increase in exercise tolerance in peripheral arterial disease, 35 and improvement in intrinsic mitochondrial efficiency.36”
From Dietary nitrate supplementation improves revascularization in chronic ischemia (2012) · cites this paper as [34]
In the citing paper
“Nitrite may form NO either via deoxyhemoglobin (deoxyHb, Fe II Hb) [80,81], deoxymyoglobin [82] or xanthine oxidoreductase (XOR)-mediated reduction [83], or via spontaneous [84] and carbonic anhydrase-facilitated disproportionation [85].”
From On the chemical biology of the nitrite/sulfide interaction (2015) · cites this paper as [83]
In the citing paper
“The extent to which RBC XOR can contribute to NO generation from nitrite will depend on the concentrations of XOR that accumulate from circulating levels [88].”
From Mechanisms of nitrite bioactivation (2014) · cites this paper as [88]
In the citing paper
“In addition to XOR mediated NO generation from nitrite occurring in heart, liver, and smooth muscle tissue, XOR has been proposed to be associated with endothelial cells and RBCs and reduce nitrite there [88].”
From Mechanisms of nitrite bioactivation (2014) · cites this paper as [88]
In the citing paper
“XOR was also demonstrated to be active on the erythrocytic membrane [25, 88].”
From Mechanisms of nitrite bioactivation (2014) · cites this paper as [88]
In the citing paper
“They also suggested that eNOS, both in the RBC and endothelial cells, played a role in vascular NO production from nitrite using a NOS blocker [88].”
From Mechanisms of nitrite bioactivation (2014) · cites this paper as [88]
In the citing paper
“Webb and colleagues showed that nitrite-mediated NO production from human left internal mammary artery was substantially (but not completely) reduced by allopurinol, an XOR inhibitor [88].”
From Mechanisms of nitrite bioactivation (2014) · cites this paper as [88]
In the citing paper
“Several studies have emphasized the importance of XOR in mediating nitrite-dependent increases in blood flow either due to XOR in the blood vessels [39, 88] or erythrocytic XOR [25, 88].”
From Mechanisms of nitrite bioactivation (2014) · cites this paper as [88]
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Metadata
- Authors
- Webb, A. J., Milsom, A. B., Rathod, K. S., Chu, W. L., Qureshi, S., Lovell, M. J., Lecomte, F. M. J., Perrett, D., Raimondo, C., Khoshbin, E., Ahmed, Z., Uppal, R., Benjamin, N., Hobbs, A. J., & Ahluwalia, A.
- Affiliations
- St Bartholomew's Hospital (GB)
- ORCID
- Venue
- Circulation Research
- Published
- 2008-10-24
- Volume
- 103
- Issue
- 9
- Pages
- 957-964
- ISSN
- 0009-7330 (ISSN-L)
- 1524-4571
- PMC
- PMC2841343
- OpenAlex
- W2158124791
- Cited by
- 203
- References
- 47
- Citations by year
- 2026: 7 · 2025: 9 · 2024: 6 · 2023: 7 · 2022: 5 · 2021: 8 · 2020: 7 · 2019: 8 · 2018: 13 · 2017: 16 · 2016: 10 · 2015: 14 · 2014: 23 · 2013: 14 · 2012: 19
- FWCI
- 8.12
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Circulation Research (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Research Portal (King's College London) (submittedVersion)
- Funders
- Wellcome Trust (067422)
- British Heart Foundation
- Topics
- Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Hemoglobin structure and function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- nitrite reduction, nitric oxide, hypoxia, xanthine oxidoreductase, erythrocytes, endothelial nitric oxide synthase, allopurinol, acidosis, L-arginine, L-NMMA, L-NAME, vasodilation, ischemia, deoxyhemoglobin, blood vessels
- MeSH terms
- Allopurinol, Animals, Aorta, Arginine, Cells, Cultured, Enzyme Inhibitors, Erythrocytes, Fluorometry, Humans, Hydrogen-Ion Concentration, Male, Nitric Oxide, Nitrite Reductases, Nitrites, Oxidation-Reduction, Xanthine Dehydrogenase, Cell Hypoxia, Rats, Wistar, omega-N-Methylarginine, NG-Nitroarginine Methyl Ester, Xanthine, Endothelial Cells, Rats, Nitric Oxide Synthase Type II, Nitric Oxide Synthase Type III
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