Nitrite supplementation reverses vascular endothelial dysfunction and large elastic artery stiffness with aging: Sodium nitrite and vascular dysfunction with aging
Open accessAbstract
We tested the hypothesis that short-term nitrite therapy reverses vascular endothelial dysfunction and large elastic artery stiffening with aging, and reduces arterial oxidative stress and inflammation. Nitrite concentrations were lower (P < 0.05) in arteries, heart, and plasma of old (26-28 month) male C57BL6 control mice, and 3 weeks of sodium nitrite (50 mg L(-1) in drinking water) restored nitrite levels to or above young (4-6 month) controls. Isolated carotid arteries of old control mice had lower acetylcholine (ACh)-induced endothelium-dependent dilation (EDD) (71.7 ± 6.1% vs. 93.0 ± 2.0%) mediated by reduced nitric oxide (NO) bioavailability (P < 0.05 vs. young), and sodium nitrite restored EDD (95.5 ± 1.6%) by increasing NO bioavailability. 4-Hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPOL), a superoxide dismutase (SOD) mimetic, apocynin, a nicotinamide adenine dinucleotide phosphate-oxidase (NADPH) inhibitor, and sepiapterin (exogenous tetrahydrobiopterin) each restored EDD to ACh in old control, but had no effect in old nitrite-supplemented mice. Old control mice had increased aortic pulse wave velocity (478 ± 16 vs. 332 ± 12 AU, P < 0.05 vs. young), which nitrite supplementation lowered (384 ± 27 AU). Nitrotyrosine, superoxide production, and expression of NADPH oxidase were ∼100-300% greater and SOD activity was ∼50% lower in old control mice (all P < 0.05 vs. young), but were ameliorated by sodium nitrite treatment. Inflammatory cytokines were markedly increased in old control mice (P < 0.05), but reduced to levels of young controls with nitrite supplementation. Short-term nitrite therapy reverses age-associated vascular endothelial dysfunction, large elastic artery stiffness, oxidative stress, and inflammation. Sodium nitrite may be a novel therapy for treating arterial aging in humans.
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Pre-clinical studies with nitrate or nitrite show the potential to protect against ischaemia-reperfusion injury and reduce arterial stiffness, inflammation and intimal thickness.
“Pre-clinical studies with nitrate or nitrite also show the potential to protect against ischaemia-reperfusion injury and reduce arterial stiffness, inflammation and intimal thickness.”
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 [150]
The Introduction reports that sodium nitrite protects against ischaemia-reperfusion injury, and the Discussion reports reduced large-artery stiffness and inflammatory markers in old mice. Our copy does not report a reduction in intimal thickness; that part of the claim is unsupported. (LLM, conf=0.87)
Experimental — accuracy not yet measuredSindler et al. found that nitrite reduced the elevated pulse wave velocity and levels of inflammatory cytokines IL-1b, IL6, INFg and TNFa.
“Thus, Sindler et al. found that nitrite reduced the elevated pulse wave velocity and levels of inflammatory cytokines IL-1b, IL6, INFg and TNFa [150].”
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 [150]
Sindler et al. found decreased plasma nitrite concentrations in old male C57BL6 control mice, depleted in arteries and heart, which were restored to youthful levels by 3 weeks' supplementation with sodium nitrite (50 mg l-1 in the drinking water).
“Sindler et al. also detected depleted concentrations in the arteries and heart, which were restored to youthful levels (4-6 month controls) by 3 weeks' supplementation with sodium nitrite (50 mg l -1 in the drinking water) [150].”
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 [150]
While the efficacy of dietary nitrate to reduce systemic blood pressure in older age has been extensively studied, relatively few studies have assessed the effects of nitrate supplementation on exercise capacity in healthy older individuals.
“While the efficacy of dietary NO 3 -to reduce systemic blood pressure in older age has been extensively studied (reviewed in 140; see also 139, 142), relatively few studies have assessed the effects of NO 3 -supplementation on exercise capacity in healthy older individuals.”
From Dietary Nitrate and Physical Performance (2018) · cites this paper as [142]
Split into 2 atoms — 0 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 2 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measuredOral supplementation with either nitrite or nitrate increases circulating nitrite levels and prevents and reverses endothelial dysfunction in a dose-dependent manner in mice and in healthy, hypertensive, and obese individuals, or in those with hypercholesterolemia and peripheral arterial disease.
“Oral supplementation with either nitrite or nitrate increases circulating nitrite levels and prevents and reverses endothelial dysfunction in a dose-dependent manner both in mice[68,69] and in healthy[70,71,97,111,112], hypertensive[113], and obese individuals[114], or in those with hypercholesterolemia[115] and peripheral arterial disease[116].”
From Enterosalivary nitrate metabolism and the microbiome: Intersection of microbial metabolism, nitric oxide and diet in cardiac and pulmonary vascular health (2017) · cites this paper as [69]
In old mice, short-term oral sodium nitrite supplementation ameliorates endothelial dysfunction, as indicated by normalization of nitric oxide-mediated endothelium-dependent dilation.
“ameliorates endothelial dysfunction, as indicated by normalization of NO-mediated endothelium-dependent dilation.”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
In old mice, short-term oral sodium nitrite supplementation reduces aortic pulse wave velocity.
“In old mice, short-term oral sodium nitrite supplementation reduces aortic pulse wave velocity, the gold-standard measure of large elastic artery stiffness”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Improvements in age-related vascular dysfunction with nitrite are mediated by reductions in oxidative stress and inflammation.
“These improvements in age-related vascular dysfunction with nitrite are mediated by reductions in oxidative stress and inflammation”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Vascular endothelial function as assessed by carotid artery endothelium-dependent dilation in response to acetylcholine was impaired in old compared with young control animals.
“vascular endothelial function as assessed by carotid artery EDD in response to acetylcholine, was impaired in old compared with young control animals (Fig. 4, top).”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Sodium nitrite treatment restored endothelium-dependent dilation in old mice by increasing nitric oxide-dependent dilation.
“Sodium nitrite treatment restored EDD in old mice by increasing NO-dependent dilation (Fig. 4, bottom).”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Staining intensity for nitrotyrosine, a cellular marker of oxidant modification of proteins, was markedly greater in aorta of old compared with young control mice.
“the staining intensity for nitrotyrosine, a cellular marker of oxidant modification of proteins, was markedly greater in aorta of old compared with young control mice (53, 171) (Fig. 5A).”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Nitrite concentrations in plasma and large elastic arteries were lower in older animals, but sodium nitrite supplementation increased nitrite concentrations in old mice up to or above those of young control animals.
“Nitrite concentrations in the plasma and large elastic arteries (aorta and carotid arteries) were lower in older animals, but sodium nitrite supplementation increased nitrite concentrations in the old mice up to or above those of young control animals (Fig. 2) (171).”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Sodium nitrite had no effect on arterial function in young mice in this study.
“sodium nitrite had no effect on arterial function in the young mice in this study (171)”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Aortic pulse wave velocity was markedly higher in old compared with young control mice.
“aortic PWV was markedly higher (i.e., the aorta was stiffer) in old compared with young control mice (Fig. 3A).”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Sodium nitrite treatment normalized aortic nitrotyrosine staining in old mice, indicating amelioration of age-associated arterial oxidative stress.
“Sodium nitrite treatment normalized aortic nitrotyrosine staining in old mice, indicating amelioration of age-associated arterial oxidative stress.”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Sodium nitrite treatment reduced aortic pulse wave velocity in old mice to well below old controls and not significantly different from that in young animals.
“Sodium nitrite treatment reduced aortic PWV in old mice to well below old controls and not significantly different from that in young animals.”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Superoxide production is much greater in aorta from old mice than from young control mice.
“Superoxide production was much greater in aorta from old compared with young control mice”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Inhibition of nitric oxide using l-NAME markedly reduced endothelium-dependent dilation in young animals while having less effect in old mice, indicating that impaired endothelium-dependent dilation in old animals was mediated by reduced nitric oxide bioavailability.
“Inhibition of NO using the NO synthase inhibitor l-NAME, markedly reduced EDD in young animals while having less effect in old mice, indicating that the impaired EDD in old animals was mediated by reduced NO bioavailability.”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Nitrite did not affect endothelium-independent dilation to sodium nitroprusside, indicating that improvements in endothelium-dependent dilation were not induced by increased vascular smooth muscle sensitivity to nitric oxide.
“Nitrite did not affect endothelium-independent dilation to sodium nitroprusside (171), indicating that the improvements in EDD were not induced by increased vascular smooth muscle sensitivity to NO.”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Expression of proinflammatory cytokines IL-1β, IL-6, interferon-γ, and TNF-α is greater in aorta of old mice than young control mice.
“expression of the proinflammatory cytokines interleukin (IL)-1β, IL-6, interferon-γ, and tumor necrosis factor-α (TNF-α) were shown to be greater in aorta of old mice than young control mice (171)”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Sepiapterin has no effect on EDD in old mice treated with sodium nitrite.
“although it had no effect on EDD in old mice treated with sodium nitrite (171)”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
SOD activity is approximately 50% lower in aorta of old compared with young control mice.
“The activity of the major antioxidant enzyme SOD was found to be ∼50% lower in aorta of old compared with young control mice”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Protein expression of NADPH oxidase is threefold greater in aorta of old mice compared with young control mice.
“Protein expression of NADPH oxidase was threefold greater in aorta of old mice compared with those of young control mice”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Inhibition of NADPH oxidase with apocynin has no effect on EDD in arteries of old animals treated with sodium nitrite.
“Inhibition of NADPH oxidase with apocynin had no effect on EDD in arteries of old animals treated with sodium nitrite”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Short-term treatment with sodium nitrite completely normalizes superoxide formation in old animals.
“short-term treatment with sodium nitrite completely normalized superoxide formation in old animals (53, 171)”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Sepiapterin restores acetylcholine-induced EDD selectively in carotid arteries of old control mice.
“Sepiapterin restored acetylcholine-induced EDD selectively in carotid arteries of old control mice”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
TEMPOL has no effect on EDD in old mice treated with sodium nitrite.
“TEMPOL had no effect on EDD in old mice treated with sodium nitrite”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Inhibition of NADPH oxidase with apocynin restores EDD in old control mice.
“but restored EDD in old control mice (171)”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Short-term treatment with sodium nitrite restores SOD activity in aorta of old mice to levels observed in young controls.
“short-term treatment with sodium nitrite restored SOD activity in aorta of old mice to levels observed in young controls”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
TEMPOL completely restores EDD in old control animals.
“but completely restored EDD in old control animals”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
NADPH oxidase protein expression is normalized after sodium nitrite treatment in old mice.
“but this was normalized after sodium nitrite treatment”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
Nitrite supplementation completely reverses age-associated arterial inflammation in old mice.
“Most importantly, nitrite supplementation completely reversed this age-associated arterial inflammation (171).”
From Inorganic nitrite supplementation for healthy arterial aging (2014) · cites this paper as [171]
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SIRT1‐mediated ERβ suppression in the endothelium contributes to vascular aging
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Does dietary nitrate say NO to cardiovascular ageing? Current evidence and implications for research
Siervo, M., Scialò, F., Shannon, O. M., Stephan, B. C. M., & Ashor, A. W. (2018) · Proceedings of The Nutrition Society · doi:10.1017/s0029665118000058 · Cited by 48 · see citing works
Ballak, D. B., Brunt, V. E., Sapinsley, Z. J., Ziemba, B. P., Richey, J. J., Zigler, M. C., Johnson, L. C., Gioscia‐Ryan, R. A., Culp‐Hill, R., Eisenmesser, E. Z., et al. (2019) · Aging Cell · doi:10.1111/acel.13074 · Cited by 47 · see citing works
Self-citationSchiffer, T. A., Lundberg, J. O., Weitzberg, E., & Carlström, M. (2020) · Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease · doi:10.1016/j.bbadis.2020.165811 · Cited by 48 · see citing works
Prasad, A. M., Morgan, D. A., Nuno, D. W., Ketsawatsomkron, P., Bair, T., Venema, A. N., Dibbern, M. E., Kutschke, W. J., Weiss, R. M., Lamping, K. G., et al. (2015) · Journal of the American Heart Association · doi:10.1161/jaha.115.001949 · Cited by 45 · see citing works
Ferguson, S. K., Holdsworth, C. T., Colburn, T. D., Wright, J. L., Craig, J. C., Fees, A. J., JONES, A. M., Allen, J. D., Musch, T. I., & Poole, D. C. (2016) · Journal of Applied Physiology · doi:10.1152/japplphysiol.00014.2016 · Cited by 45 · see citing works
Inorganic Nitrite Supplementation Improves Endothelial Function With Aging
Rossman, M. J., Gioscia‐Ryan, R. A., Santos‐Parker, J. R., Ziemba, B. P., Lubieniecki, K. L., Johnson, L. C., Poliektov, N. E., Bispham, N. Z., Woodward, K. A., Nagy, E., et al. (2021) · Hypertension · doi:10.1161/hypertensionaha.120.16175 · Cited by 46 · see citing works
Self-citationA “green” diet-based approach to cardiovascular health? Is inorganic nitrate the answer?
Rathod, K. S., Velmurugan, S., & Ahluwalia, A. (2016) · Molecular Nutrition & Food Research · doi:10.1002/mnfr.201500313 · Cited by 42 · see citing works
Hughes, W. E., Ueda, K., Treichler, D. P., & Casey, D. P. (2016) · Nitric Oxide · doi:10.1016/j.niox.2016.06.007 · Cited by 39 · see citing works
Clayton, Z. S., Brunt, V. E., Hutton, D. A., Casso, A. G., Ziemba, B. P., Melov, S., Campisi, J., & Seals, D. R. (2021) · Hypertension · doi:10.1161/hypertensionaha.120.16759 · Cited by 40 · see citing works
Self-citationThe relationship between plasma and salivary NOx
Clodfelter, W. H., Basu, S., Bolden, C., Dos Santos, P. C., King, S. B., & Kim-Shapiro, D. B. (2015) · Nitric Oxide · doi:10.1016/j.niox.2015.04.003 · Cited by 39 · see citing works
Ling, W. C., Mustafa, M. R., Vanhoutte, P. M., & Murugan, D. D. (2017) · Vascular Pharmacology · doi:10.1016/j.vph.2017.05.003 · Cited by 40 · see citing works
Sex differences in vascular aging and impact of GPER deletion
Ogola, B. O., Abshire, C., Visniauskas, B., Kiley, J. X., Horton, A. C., Clark, G. L., Kilanowski‐Doroh, I., Diaz, Z. T., Bicego, A. N., McNally, A. B., et al. (2022) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00238.2022 · Cited by 39 · see citing works
- Authors
- Sindler, A. L., Fleenor, B. S., Calvert, J. W., Marshall, K. D., Zigler, M. L., Lefer, D. J., & Seals, D. R.
- Affiliations
- University of Colorado Boulder (US)
- Emory University (US)
- Venue
- Aging Cell
- Publisher
- Wiley
- Published
- 2011-03-31
- Volume
- 10
- Issue
- 3
- Pages
- 429-437
- ISSN
- 1474-9718 (ISSN-L)
- PMC
- PMC3094511
- OpenAlex
- W1559904992
- Cited by
- 218
- References
- 56
- Citations by year
- 2026: 3 · 2025: 5 · 2024: 11 · 2023: 8 · 2022: 12 · 2021: 17 · 2020: 16 · 2019: 15 · 2018: 14 · 2017: 17 · 2016: 19 · 2015: 23 · 2014: 22 · 2013: 11 · 2012: 17
- FWCI
- 12.54
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Aging Cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute on Aging (AG-000279, R37AG013038, R01AG013038, AG-013038, T32AG000279)
- National Heart, Lung, and Blood Institute (R01HL098481, HL007822, R01HL093579, T32HL007822, HL092141, R01HL092141, HL093579)
- Topics
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiovascular Health and Disease Prevention
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiovascular Function and Risk Factors
- Keywords
- nitrite supplementation, sodium nitrite, endothelial dysfunction, arterial stiffness, aging, nitric oxide bioavailability, oxidative stress, inflammation, NADPH oxidase, superoxide, tempol, apocynin, sepiapterin, pulse wave velocity, carotid artery, nitrotyrosine, inflammatory cytokines, arterial aging
- MeSH terms
- Acetophenones, Acetylcholine, Administration, Oral, Aging, Animals, Biopterins, Carotid Arteries, Cyclic N-Oxides, Elasticity, Endothelium, Vascular, Humans, Male, Myography, Nitric Oxide, Sodium Nitrite, Spin Labels, Superoxide Dismutase, Vasodilation, Vasodilator Agents, Cytokines, Oxidative Stress, NADPH Oxidases, Mice
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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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