Subjects

Abstract

Nitrite anions comprise the largest vascular storage pool of nitric oxide (NO), provided that physiological mechanisms exist to reduce nitrite to NO. We evaluated the vasodilator properties and mechanisms for bioactivation of nitrite in the human forearm. Nitrite infusions of 36 and 0.36 μmol/min into the forearm brachial artery resulted in supra- and near-physiologic intravascular nitrite concentrations, respectively, and increased forearm blood flow before and during exercise, with or without NO synthase inhibition. Nitrite infusions were associated with rapid formation of erythrocyte iron-nitrosylated hemoglobin and, to a lesser extent, S-nitroso-hemoglobin. NO-modified hemoglobin formation was inversely proportional to oxyhemoglobin saturation. Vasodilation of rat aortic rings and formation of both NO gas and NO-modified hemoglobin resulted from the nitrite reductase activity of deoxyhemoglobin and deoxygenated erythrocytes. This finding links tissue hypoxia, hemoglobin allostery and nitrite bioactivation. These results suggest that nitrite represents a major bioavailable pool of NO, and describe a new physiological function for hemoglobin as a nitrite reductase, potentially contributing to hypoxic vasodilation.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    & Requirement of thiols for activation of coronary arterial guanylate cyclase by glyceryl trinitrate and sodium nitrite: possible involvement of S-nitrosothiols. Biochim. Biophys. Acta. 631, 221–231 (1980).

  2. 2.

    et al. Mechanism of vascular smooth muscle relaxation by organic nitrates, nitrites, nitroprusside and nitric oxide: evidence for the involvement of S-nitrosothiols as active intermediates. J. Pharmacol. Exp. Ther. 218, 739–749 (1981).

  3. 3.

    , & A comparison of the effects of hydrallazine, diazoxide, sodium nitrite and sodium nitroprusside on human isolated arteries and veins. Br. J. Clin. Pharmacol. 11, 57–61 (1981).

  4. 4.

    , , , & Relationship between cyclic guanosine 3':5′-monophosphate formation and relaxation of coronary arterial smooth muscle by glyceryl trinitrate, nitroprusside, nitrite and nitric oxide: effects of methylene blue and methemoglobin. J. Pharmacol. Exp. Ther. 219, 181–186 (1981).

  5. 5.

    & Interactions of light and sodium nitrite in producing relaxation of rabbit aorta. J. Pharmacol. Exp. Ther. 248, 687–695 (1989).

  6. 6.

    et al. Exogenous GTP enhances the effects of sodium nitrite on cyclic GMP accumulation, vascular smooth muscle relaxation and platelet aggregation. Pharmacol. Toxicol. 68, 60–63 (1991).

  7. 7.

    , , , & Chemical nature of nitric oxide storage forms in rat vascular tissue. Proc. Natl. Acad. Sci. USA 100, 336–341 (2003).

  8. 8.

    et al. Role of circulating nitrite and S-nitrosohemoglobin in the regulation of regional blood flow in humans. Proc. Natl. Acad. Sci. USA 97, 11482–11487 (2000).

  9. 9.

    et al. NO adducts in mammalian red blood cells: too much or too little? Nat. Med. 9, 481–483 (2003).

  10. 10.

    , , & Concomitant presence of N-nitroso and S-nitroso proteins in human plasma. Free Radic. Biol. Med. 33, 1590–1596 (2002).

  11. 11.

    et al. Evidence for in vivo transport of bioactive nitric oxide in human plasma. J. Clin. Invest. 109, 1241–1248 (2002).

  12. 12.

    , & NO solutions? J. Clin. Invest. 109, 1149–1151 (2002).

  13. 13.

    , & Xanthine oxidase can generate nitric oxide from nitrate in ischaemia. Biochem. Soc. Trans. 25, 528S (1997).

  14. 14.

    et al. Xanthine oxidoreductase catalyses the reduction of nitrates and nitrite to nitric oxide under hypoxic conditions. FEBS Lett. 427, 225–228 (1998).

  15. 15.

    et al. Reduction of nitrite to nitric oxide catalyzed by xanthine oxidoreductase. J. Biol. Chem. 275, 7757–7763 (2000).

  16. 16.

    et al. Generation of nitric oxide by a nitrite reductase activity of xanthine oxidase: a potential pathway for nitric oxide formation in the absence of nitric oxide synthase activity. Biochem. Biophys. Res. Commun. 249, 767–772 (1998).

  17. 17.

    , , & Characterization of the magnitude and kinetics of xanthine oxidase-catalyzed nitrite reduction. Evaluation of its role in nitric oxide generation in anoxic tissues. J. Biol. Chem. 276, 24482–24489 (2001).

  18. 18.

    , , & Characterization of the magnitude and kinetics of xanthine oxidase-catalyzed nitrate reduction: evaluation of its role in nitrite and nitric oxide generation in anoxic tissues. Biochemistry 42, 1150–1159 (2003).

  19. 19.

    , , & Enzyme-independent formation of nitric oxide in biological tissues. Nat. Med. 1, 804–809 (1995).

  20. 20.

    , & Non-enzymatic nitric oxide synthesis in biological systems. Biochim. Biophys. Acta 1411, 250–262 (1999).

  21. 21.

    , & Evaluation of the magnitude and rate of nitric oxide production from nitrite in biological systems. Arch. Biochem. Biophys. 357, 1–7 (1998).

  22. 22.

    et al. Nitrite-derived nitric oxide: a possible mediator of 'acidic-metabolic' vasodilation. Acta Physiol. Scand. 171, 9–16 (2001).

  23. 23.

    et al. Circulating nitrite anions are a directly acting vasodilator and are donors for nitric oxide. Clin. Sci. (Lond.) 102, 77–83 (2002).

  24. 24.

    , , , & Mechanisms of nitric oxide generation from nitroglycerin and endogenous sources during hypoxia in vivo. Br. J. Pharmacol. 135, 373–382 (2002).

  25. 25.

    et al. Plasma nitrite rather than nitrate reflects regional endothelial nitric oxide synthase activity but lacks intrinsic vasodilator action. Proc. Natl. Acad. Sci. USA 98, 12814–12819 (2001).

  26. 26.

    et al. Different plasma levels of nitric oxide in arterial and venous blood. Clin. Physiol. 19, 440–442 (1999).

  27. 27.

    et al. Inhaled NO as a viable antiadhesive therapy for ischemia/reperfusion injury of distal microvascular beds. J. Clin. Invest. 101, 2497–2505 (1998).

  28. 28.

    et al. Nitric oxide in the human respiratory cycle. Nat. Med. 3, 3 (2002).

  29. 29.

    et al. Effects of inhaled nitric oxide on regional blood flow are consistent with intravascular nitric oxide delivery. J. Clin. Invest. 108, 279–287 (2001).

  30. 30.

    et al. S-nitrosohemoglobin is unstable in the reductive red cell environment and lacks O2/NO-linked allosteric function. J. Biol. Chem. 21, 21 (2002).

  31. 31.

    , , & Nitric oxide's reactions with hemoglobin: a view through the SNO-storm. Nat. Med. 9, 496–500 (2003).

  32. 32.

    & Hemoglobin and the paracrine and endocrine functions of nitric oxide. N. Engl. J. Med. 348, 1483–1485 (2003).

  33. 33.

    , , , & Kinetics and mechanism of the oxidation of human deoxyhemoglobin by nitrites. J. Biol. Chem. 256, 12393–12398 (1981).

  34. 34.

    et al. Routes to S-nitroso-hemoglobin formation with heme redox and preferential reactivity in the beta subunits. Proc. Natl. Acad. Sci. USA 100, 461–466 (2003).

  35. 35.

    et al. Measurements of nitric oxide on the heme iron and β-93 thiol of human hemoglobin during cycles of oxygenation and deoxygenation. Proc. Natl. Acad. Sci. USA 100, 11303–11308 (2003).

  36. 36.

    , & Nitrite catalyzes reductive nitrosylation of the water-soluble ferri-heme model Fe(III)(TPPS) to Fe(II)(TPPS)(NO). Inorg. Chem. 42, 2–4 (2003).

  37. 37.

    & Generation of superoxide and hydrogen peroxide during interaction of nitrite with human hemoglobin. Acta Med. Okayama 35, 173–178 (1981).

  38. 38.

    , & Mechanism of autocatalytic oxidation of oxyhemoglobin by nitrite. An intermediate detected by electron spin resonance. Biochim. Biophys. Acta 702, 237–241 (1982).

  39. 39.

    & Mechanism of autocatalytic oxidation of oxyhemoglobin by nitrite. Environ. Health Perspect. 73, 147–151 (1987).

  40. 40.

    , , & S-nitrosohaemoglobin: a dynamic activity of blood involved in vascular control. Nature 380, 221–226 (1996).

  41. 41.

    et al. Blood flow regulation by S-nitrosohemoglobin in the physiological oxygen gradient. Science 276, 2034–2037 (1997).

  42. 42.

    et al. Modulation of nitric oxide bioavailability by erythrocytes. Proc. Natl. Acad. Sci. USA 98, 11771–11776 (2001).

  43. 43.

    et al. Cell-free hemoglobin limits nitric oxide bioavailability in sickle-cell disease. Nat. Med. 8, 1383–1389 (2002).

  44. 44.

    , , & Role of endothelium-derived nitric oxide in the abnormal endothelium- dependent vascular relaxation of patients with essential hypertension. Circulation 87, 1468–1474 (1993).

  45. 45.

    , , & Methodologies for the sensitive and specific measurement of S-nitrosothiols, iron-nitrosyls, and nitrite in biological samples. Free Radic. Res. 37, 1–10 (2003).

  46. 46.

    , & Vasoactivity of S-nitrosohemoglobin: role of oxygen, heme, and NO oxidation states. Blood 101, 4408–4415 (2003).

Download references

Acknowledgements

This work was supported by Clinical Center and National Heart, Lung and Blood Institute intramural funds (R.O.C. and M.T.G.), National Institutes of Health grant HL58091 (D.B.K.-S.), RO1HL70146 (R.P.P.) and Medical Scientist Training Program T32GM08361. We thank V. Annavajjhala for helpful laboratory assistance.

Author information

Author notes

    • Richard O Cannon III
    •  & Mark T Gladwin

    These authors contributed equally to this work.

Affiliations

  1. Cardiovascular Branch, National Heart, Lung and Blood Institute, National Institutes of Health, 10 Center Drive, Building 10, Room 7B15 Bethesda, Maryland 20892, USA.

    • Kenyatta Cosby
    • , Gloria Zalos
    •  & Richard O Cannon III
  2. Critical Care Medicine Department, Warren G. Magnuson Clinical Center, National Institutes of Health, 10 Center Drive, Building 10, Room 7D43 Bethesda, Maryland 20892, USA.

    • Kristine S Partovi
    • , Christopher D Reiter
    • , Sabrina Martyr
    • , Benjamin K Yang
    •  & Mark T Gladwin
  3. Department of Pathology, Center for Free Radical Biology, Biomedical Research Building II, Room 307, 901 19th Street South, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.

    • Jack H Crawford
    •  & Rakesh P Patel
  4. Laboratory of Chemical Biology, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, 10 Center Drive, Building 10, Room 9N307, Bethesda, Maryland 20892, USA.

    • Christopher D Reiter
    • , Sabrina Martyr
    • , Alan N Schechter
    •  & Mark T Gladwin
  5. Office of Biostatistics Research, National Heart, Lung and Blood Institute, National Institutes of Health, 6701 Rockledge Drive, Bethesda, Maryland 20892, USA.

    • Myron A Waclawiw
  6. Department of Physics, Wake Forest University, Winston-Salem, North Carolina 27109-7507, USA.

    • Xiuli Xu
    • , Howard Shields
    •  & Daniel B Kim-Shapiro
  7. Department of Biomedical Engineering, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27157-1022, USA.

    • Kris T Huang
    •  & Daniel B Kim-Shapiro

Authors

  1. Search for Kenyatta Cosby in:

  2. Search for Kristine S Partovi in:

  3. Search for Jack H Crawford in:

  4. Search for Rakesh P Patel in:

  5. Search for Christopher D Reiter in:

  6. Search for Sabrina Martyr in:

  7. Search for Benjamin K Yang in:

  8. Search for Myron A Waclawiw in:

  9. Search for Gloria Zalos in:

  10. Search for Xiuli Xu in:

  11. Search for Kris T Huang in:

  12. Search for Howard Shields in:

  13. Search for Daniel B Kim-Shapiro in:

  14. Search for Alan N Schechter in:

  15. Search for Richard O Cannon in:

  16. Search for Mark T Gladwin in:

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Richard O Cannon III or Mark T Gladwin.

Supplementary information

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nm954