Antiplatelet effects of dietary nitrate in healthy volunteers: Involvement of cGMP and influence of sex
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Ingestion of vegetables rich in inorganic nitrate has emerged as an effective method, via the formation of a nitrite intermediate, for acutely elevating vascular NO levels. As such a number of beneficial effects of dietary nitrate ingestion have been demonstrated including the suggestion that platelet reactivity is reduced. In this study we investigated whether inorganic nitrate supplementation might also reduce platelet reactivity in healthy volunteers and have determined the mechanisms involved in the effects seen. We conducted two randomised crossover studies each in 24 (12 of each sex) healthy subjects assessing the acute effects of dietary nitrate (250 ml beetroot juice) or potassium nitrate capsules (KNO3, 8 mmol) vs placebo control on platelet reactivity. Inorganic nitrate ingested either from a dietary source or via supplementation raised circulating nitrate and nitrite levels in both sexes and attenuated ex vivo platelet aggregation responses to ADP and, albeit to a lesser extent, collagen but not epinephrine in male but not female volunteers. These inhibitory effects were associated with a reduced platelet P-selectin expression and elevated platelet cGMP levels. In addition, we show that nitrite reduction to NO occurs at the level of the erythrocyte and not the platelet. In summary, our results demonstrate that inorganic nitrate ingestion, whether via the diet or through supplementation, causes a modest decrease in platelet reactivity in healthy males but not females. Our studies provide strong support for further clinical trials investigating the potential of dietary nitrate as an adjunct to current antiplatelet therapies to prevent atherothrombotic complications. Moreover, our observations highlight a previously unknown sexual dimorphism in platelet reactivity to NO and intimate a greater dependence of males on the NO-soluble guanylate cyclase pathway in limiting thrombotic potential.
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When NOS binds BH2 instead of BH4, not only is NO synthesis completely abolished but the protein also becomes an oxidase, generating superoxide instead of NO.
“Thus, not only is the NO synthesis completely abolished but the protein also becomes an oxidase, generating superoxide instead of NO (191, 972, 1007) (Table 6).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [972]
When NOS binds BH2 instead of BH4, the ferrous/oxy heme cannot hydroxylate l-arginine to NG-hydroxy-l-arginine, NO synthesis is blocked, and the oxygen is eventually oxidized to superoxide.
“Mechanistically, when NOS binds BH2 instead of BH4, the ferrous/oxy heme cannot hydroxylate l-arginine to NG-hydroxy-l-arginine, NO synthesis is blocked, and the oxygen is eventually oxidized to superoxide.”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [972]
The entero-salivary circulation is intact and functioning normally in hypertensive patients.
“This is similar to the ratios identified in healthy volunteers following single dose nitrate supplementation28, 29 suggesting that the entero-salivary circulation is intact and functioning normally in hypertensive patients.”
From Dietary Nitrate Provides Sustained Blood Pressure Lowering in Hypertensive Patients: A Randomized, Phase 2, Double-Blind, Placebo-Controlled Study (2015) · cites this paper as [29]
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- Authors
- Velmurugan, S., Kapil, V., Ghosh, S. M., Davies, S., McKnight, A., Aboud, Z., Khambata, R. S., Webb, A. J., Poole, A., & Ahluwalia, A.
- Affiliations
- Queen Mary University of London (GB)
- University of Bristol (GB)
- Corresponding author
- Ahluwalia, Amrita
- Venue
- Free Radical Biology and Medicine
- Publisher
- Elsevier BV
- Published
- 2013-12
- Volume
- 65
- Pages
- 1521-1532
- ISSN
- 0891-5849 (ISSN-L)
- PMC
- PMC3878381
- OpenAlex
- W2028916136
- Cited by
- 114
- References
- 60
- Citations by year
- 2026: 3 · 2025: 4 · 2024: 4 · 2023: 6 · 2022: 7 · 2021: 7 · 2020: 5 · 2019: 8 · 2018: 15 · 2017: 9 · 2016: 18 · 2015: 10 · 2014: 15 · 2013: 2
- FWCI
- 10.42
- Type
- journal article
- Language
- en
- Update notices
- erratum (2015-07-01)
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Free Radical Biology and Medicine (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Research Portal (King's College London) (submittedVersion, other-oa)
- Funders
- British Heart Foundation (FS/11/41/28749)
- Medical Research Council
- Topics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Health Sciences › Medicine › Hematology › Platelet Disorders and Treatments
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Antiplatelet Therapy and Cardiovascular Diseases
- Keywords
- dietary nitrate, inorganic nitrate supplementation, platelet reactivity, platelet aggregation, nitric oxide, cyclic GMP, soluble guanylate cyclase, nitrite reduction, erythrocytes, P-selectin, sex differences, beetroot juice, potassium nitrate, atherothrombosis, antiplatelet therapy
- MeSH terms
- Adolescent, Adult, Blood Platelets, Cardiovascular Diseases, Collagen, Diet, Epinephrine, Erythrocytes, Female, Cyclic GMP, Guanylate Cyclase, Humans, Male, Middle Aged, Nitrates, Nitric Oxide, Platelet Aggregation, Platelet Aggregation Inhibitors, Sex Factors, Vegetables, Potassium Compounds, Cross-Over Studies, P-Selectin, Dietary Supplements, Beta vulgaris, Young Adult
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