Skip to main content

Nitrite is a cGMP generator in isolated platelets

cGMP is generated in blood platelets following activation of soluble guanylate cyclase (sGC) by nitric oxide (NO). Once synthesised, cGMP activates cGMP-dependent protein kinase (PKG) and causes an increase in intracellular cAMP concentration by inhibiting phosphodiesterase 3 (PDE3), responsible for its degradation. PKG and the cAMP-dependent protein kinase (PKA) phosphorylate a number of substrates with inhibitory effects on platelet function [1]. While NO is mainly generated by endothelial NO synthase (eNOS) in the endothelium, reduction of nitrite constitutes an alternative vascular source of NO during hypoxic and acidic conditions. Nitrite originates from reduction of dietary nitrate, found in foods such as beetroot and green leafy vegetables. Nitrite in the circulation can be converted to NO by several nitrite reductases in blood and tissues [2], and triggers vasodilatation [3] thus lowering blood pressure [4]. It has also been demonstrated that when eNOS activity is impaired, NO generated through the nitrate/nitrite pathway exerts negative effects on platelet function [5].

Whilst deoxyhaemoglobin is a candidate nitrite-reductase in blood [6], we aimed to characterise the effects of nitrite in isolated platelets. As such, we used a preparation of washed platelets and supraphysiological concentrations of nitrite in vitro and characterised nitrite-dependent cGMP generation and inhibition of platelet function.

Our data show that nitrite at high concentrations (1mM) is a powerful generator of cGMP in platelets in the absence of extracellular reductases. While cGMP accumulation is inhibited by sGC inhibitor ODQ, NO scavengers have a partial effect. This seems to indicate that nitrite acts through both NO-dependent and independent mechanisms. Accordingly, phosphorylation of the the cAMP and cGMP-dependent substrate VASP is strongly increased and, similarly to cGMP generation, is completely dependent on sGC activity but not NO. The increase of cGMP induced by high concentrations of nitrite inhibits platelet function as measured by platelet aggregation and secretion. However, whilst being dependent on sGC activity, the effect of nitrite on aggregation is not dependent on NO.generation. Lower concentrations of nitrite synergise with inhibition of PDEs, in particular PDE5 responsible for cGMP degradation, to trigger detectable VASP phosphorylation and inhibition of aggregation.

Nitrite triggers generation of cGMP in platelets and exerts inhibitory effects independently of extracellular reductases and other blood cells. Nitrite acts partly through reduction to NO and partly through an uncharacterised direct effect on sGC.


  1. Smolenski A: Novel roles of cAMP/cGMP-dependent signaling in platelets. J Thromb Haemost. 2012, 10 (2): 167-176. 10.1111/j.1538-7836.2011.04576.x.

    Article  CAS  PubMed  Google Scholar 

  2. Lundberg JO, Carlstrom M, Larsen FJ, Weitzberg E: Roles of dietary inorganic nitrate in cardiovascular health and disease. Cardiovasc Res. 2011, 89 (3): 525-532. 10.1093/cvr/cvq325.

    Article  CAS  PubMed  Google Scholar 

  3. Bailey JC, Feelisch M, Horowitz JD, Frenneaux MP, Madhani M: Pharmacology and therapeutic role of inorganic nitrite and nitrate in vasodilatation. Pharmacol Ther. 2014, 144 (3): 303-320. 10.1016/j.pharmthera.2014.06.009.

    Article  CAS  PubMed  Google Scholar 

  4. Kapil V, Milsom AB, Okorie M, Maleki-Toyserkani S, Akram F, Rehman F, et al: Inorganic nitrate supplementation lowers blood pressure in humans: role for nitrite-derived NO. Hypertension. 2010, 56 (2): 274-281. 10.1161/HYPERTENSIONAHA.110.153536.

    Article  CAS  PubMed  Google Scholar 

  5. Apostoli GL, Solomon A, Smallwood MJ, Winyard PG, Emerson M: Role of inorganic nitrate and nitrite in driving nitric oxide–cGMP-mediated inhibition of platelet aggregation in vitro and in vivo. J Thromb Haemost. 2014, 12 (11): 1880-1889. 10.1111/jth.12711.

    Article  CAS  PubMed  Google Scholar 

  6. Corti P, Tejero J, Gladwin MT: Evidence mounts that red cells and deoxyhemoglobin can reduce nitrite to bioactive NO to mediate intravascular endocrine NO signaling: commentary on “Anti-platelet effects of dietary nitrate in healthy volunteers: involvement of cGMP and influence of sex”. Free Radic Biol Med. 2013, 65: 1518-1520.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to Melanie Madhani.

Rights and permissions

Open Access  This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.

The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this licence, visit

The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Borgognone, A., Loka, T., Chimen, M. et al. Nitrite is a cGMP generator in isolated platelets. BMC Pharmacol Toxicol 16 (Suppl 1), A65 (2015).

Download citation

  • Published:

  • DOI: