Archives of Toxicology

, Volume 80, Issue 12, pp 811–816 | Cite as

Clinical evidence for lead-induced inhibition of nitric oxide formation

  • Fernando BarbosaJr
  • Jonas T. C. Sertorio
  • Raquel F. Gerlach
  • Jose E. Tanus-Santos
Inorganic Compounds


Lead exposure has been associated with increased cardiovascular risk, which may result, at least in part, from lead-induced increases in oxidative stress and depressed nitric oxide (NO) availability. However, no previous clinical study has examined whether lead exposure is associated with significant effects on biomarkers of NO activity (plasma nitrites, nitrates, and cyclic guanosine 3′,5′-monophosphate; cGMP). We investigated whether there is an association between the circulating concentrations of nitrites, nitrates, and cGMP and the concentrations of lead in whole blood (B-Pb) or plasma (P-Pb) from 62 lead-exposed subjects (30 men and 32 women). P-Pb was determined by inductively coupled plasma mass spectrometry and B-Pb by graphite furnace atomic absorption spectrometry. Plasma nitrite and nitrate concentrations were measured using an ozone-based chemiluminescence assay. Plasma cGMP concentrations were measured using a commercial enzyme immunoassay. We found a negative correlation between plasma nitrite and B-Pb concentrations (= −0.358; = 0.004), and between plasma nitrite and P-Pb concentrations (= −0.264; = 0.038), thus suggesting increased inhibition of NO formation with increasing B-Pb or P-Pb concentrations. However, no significant correlations were found between plasma nitrate or cGMP and B-Pb or P-Pb concentrations (all > 0.05). These findings suggest a significant inhibitory effect of lead exposure on NO formation and provide clinical evidence for a biological mechanism possibly involved the association between lead exposure and increased cardiovascular risk.


Cyclic GMP Lead toxicology Nitric oxide Nitrates Nitrites Plasma lead Whole blood lead 


  1. ATSDR (1999) Toxicological Profile for Lead. US Department of Health and Human Services, Public Health Services, AtlantaGoogle Scholar
  2. Barbosa F Jr, Tanus-Santos JE, Gerlach RF, Parsons PJ (2005) Current needs and limitations on the use of biomarkers of internal dose to diagnose lead exposure. Environ Health Perspect 113:1669–1674PubMedCrossRefGoogle Scholar
  3. Barbosa F Jr, Sandrim VC, Uzuelli JA, Gerlach RF, Tanus-Santos JE (2006) eNOS genotype-dependent correlation between whole blood lead and plasma nitric oxide products concentrations. Nitric Oxide 14:58–64PubMedCrossRefGoogle Scholar
  4. Baylis C, Vallance P (1998) Measurement of nitrite and nitrate levels in plasma and urine—what does this measure tell us about the activity of the endogenous nitric oxide system? Curr Opin Nephrol Hypertens 7:59–62PubMedGoogle Scholar
  5. Blazka ME, Harry GJ, Luster MI (1994) Effect of lead acetate on nitrite production by murine brain endothelial cell cultures. Toxicol Appl Pharmacol 126:191–194PubMedCrossRefGoogle Scholar
  6. Carmignani M, Volpe AR, Boscolo P, Qiao N, Di Gioacchino M, Grilli A, Felaco M (2000) Catcholamine and nitric oxide systems as targets of chronic lead exposure in inducing selective functional impairment. Life Sci 68:401–415PubMedCrossRefGoogle Scholar
  7. Cooke JP, Dzau VJ (1997) Nitric oxide synthase: role in the genesis of vascular disease. Annu Rev Med 48:489–509PubMedCrossRefGoogle Scholar
  8. Ellis G, Adatia I, Yazdanpanah M, Makela SK (1998) Nitrite and nitrate analyses: a clinical biochemistry perspective. Clin Biochem 31:195–220PubMedCrossRefGoogle Scholar
  9. Garcia-Arenas G, Claudio L, Perez-Severiano F, Rios C (1999) Lead acetate exposure inhibits nitric oxide synthase activity in capillary and synaptosomal fractions of mouse brain. Toxicol Sci 50:244–248PubMedCrossRefGoogle Scholar
  10. Gulson BL, Jameson CW, Mahaffey KR, Mizon KJ, Korsch MJ, Vimpani G (1997) Pregnancy increases mobilization of lead from maternal skeleton. J Lab Clin Med 130:51–62PubMedCrossRefGoogle Scholar
  11. Gulson BL, Mahaffey KR, Jameson CW, Mizon KJ, Korsch MJ, Cameron MA, Eisman JA (1998) Mobilization of lead from the skeleton during the postnatal period is larger than during pregnancy. J Lab Clin Med 131:324–329PubMedCrossRefGoogle Scholar
  12. Gulson BL, Pounds JG, Mushak P, Thomas BJ, Gray B, Korsch MJ (1999) Estimation of cumulative lead releases (lead flux) from the maternal skeleton during pregnancy and lactation. J Lab Clin Med 134:631–640PubMedCrossRefGoogle Scholar
  13. Gurer H, Ercal N (2000) Can antioxidants be beneficial in the treatment of lead poisoning? Free Radic Biol Med 29:927–945PubMedCrossRefGoogle Scholar
  14. Gurer-Orhan H, Sabir HU, Ozgunes H (2004) Correlation between clinical indicators of lead poisoning and oxidative stress parameters in controls and lead-exposed workers. Toxicology 195:147–154PubMedCrossRefGoogle Scholar
  15. Hibbs JB Jr, Westenfelder C, Taintor R, Vavrin Z, Kablitz C, Baranowski RL, Ward JH, Menlove RL, McMurry MP, Kushner JP et al (1992) Evidence for cytokine-inducible nitric oxide synthesis from l-arginine in patients receiving interleukin-2 therapy. J Clin Invest 89:867–877PubMedCrossRefGoogle Scholar
  16. Hirata Y, Ishii M, Matsuoka H, Sugimoto T, Iizuka M, Uchida Y, Serizawa T, Sato H, Kohmoto O, Mochizuki T et al (1987) Plasma concentrations of alpha-human atrial natriuretic polypeptide and cyclic GMP in patients with heart disease. Am Heart J 113:1463–1469PubMedCrossRefGoogle Scholar
  17. Ignarro LJ, Buga GM, Wood KS, Byrns RE, Chaudhuri G (1987) Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proc Natl Acad Sci USA 84:9265–9269PubMedCrossRefGoogle Scholar
  18. Kelm M, Preik-Steinhoff H, Preik M, Strauer BE (1999) Serum nitrite sensitively reflects endothelial NO formation in human forearm vasculature: evidence for biochemical assessment of the endothelial l-arginine-NO pathway. Cardiovasc Res 41:765–772PubMedCrossRefGoogle Scholar
  19. Kielstein JT, Impraim B, Simmel S, Bode-Boger SM, Tsikas D, Frolich JC, Hoeper MM, Haller H, Fliser D (2004) Cardiovascular effects of systemic nitric oxide synthase inhibition with asymmetrical dimethylarginine in humans. Circulation 109:172–177PubMedCrossRefGoogle Scholar
  20. Kleinbongard P, Dejam A, Lauer T, Rassaf T, Schindler A, Picker O, Scheeren T, Godecke A, Schrader J, Schulz R, Heusch G, Schaub GA, Bryan NS, Feelisch M, Kelm M (2003) Plasma nitrite reflects constitutive nitric oxide synthase activity in mammals. Free Radic Biol Med 35:790–796PubMedCrossRefGoogle Scholar
  21. Kleinbongard P, Dejam A, Lauer T, Jax T, Kerber S, Gharini P, Balzer J, Zotz RB, Scharf RE, Willers R, Schechter AN, Feelisch M, Kelm M (2006) Plasma nitrite concentrations reflect the degree of endothelial dysfunction in humans. Free Radic Biol Med 40:295–302PubMedCrossRefGoogle Scholar
  22. Lauer T, Preik M, Rassaf T, Strauer BE, Deussen A, Feelisch M, Kelm M (2001) Plasma nitrite rather than nitrate reflects regional endothelial nitric oxide synthase activity but lacks intrinsic vasodilator action. Proc Natl Acad Sci USA 98:12814–12819PubMedCrossRefGoogle Scholar
  23. Metzger IF, Souza-Costa DC, Marroni AS, Nagassaki S, Desta Z, Flockhart DA, Tanus-Santos JE (2005) Endothelial nitric oxide synthase gene haplotypes associated with circulating concentrations of nitric oxide products in healthy men. Pharmacogenet Genomics 15:565–570PubMedGoogle Scholar
  24. Moller L, Kristensen TS (1992) Blood lead as a cardiovascular risk factor. Am J Epidemiol 136:1091–1100PubMedGoogle Scholar
  25. Montenegro MF, Barbosa F Jr, Sandrim VC, Gerlach RF, Tanus-Santos JE (2006) A polymorphism in the delta-aminolevulinic acid dehydratase gene modifies plasma/whole blood lead ratio. Arch Toxicol (in press). DOI 10.1007/s00204-005-0056-yGoogle Scholar
  26. Nagassaki S, Metzger IF, Souza-Costa DC, Marroni AS, Uzuelli JA, Tanus-Santos JE (2005) eNOS genotype is without effect on circulating nitrite/nitrate level in healthy male population. Thromb Res 115:375–379PubMedCrossRefGoogle Scholar
  27. Navas-Acien A, Selvin E, Sharrett AR, Calderon-Aranda E, Silbergeld E, Guallar E (2004) Lead, cadmium, smoking, and increased risk of peripheral arterial disease. Circulation 109:3196–3201PubMedCrossRefGoogle Scholar
  28. Ni Z, Hou S, Barton CH, Vaziri ND (2004) Lead exposure raises superoxide and hydrogen peroxide in human endothelial and vascular smooth muscle cells. Kidney Int 66:2329–2336PubMedCrossRefGoogle Scholar
  29. Palmer RM, Ferrige AG, Moncada S (1987) Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature 327:524–526PubMedCrossRefGoogle Scholar
  30. Pirkle JL, Schwartz J, Landis JR, Harlan WR (1985) The relationship between blood lead levels and blood pressure and its cardiovascular risk implications. Am J Epidemiol 121:246–258PubMedGoogle Scholar
  31. Rosselli M, Imthurn B, Keller PJ, Jackson EK, Dubey RK (1995) Circulating nitric oxide (nitrite/nitrate) levels in postmenopausal women substituted with 17 beta-estradiol and norethisterone acetate. A two-year follow-up study. Hypertension 25:848–853PubMedGoogle Scholar
  32. Schutz A, Bergdahl IA, Ekholm A, Skerfving S (1996) Measurement by ICP-MS of lead in plasma and whole blood of lead workers and controls. Occup Environ Med 53:736–740PubMedCrossRefGoogle Scholar
  33. Schwartz J (1995) Lead, blood pressure, and cardiovascular disease in men. Arch Environ Health 50:31–37PubMedCrossRefGoogle Scholar
  34. Silbergeld EK, Schwartz J, Mahaffey K (1988) Lead and osteoporosis: mobilization of lead from bone in postmenopausal women. Environ Res 47:79–94PubMedCrossRefGoogle Scholar
  35. Souza-Silva AR, Dias-Junior CA, Uzuelli JA, Moreno H Jr, Evora PR, Tanus-Santos JE (2005) Hemodynamic effects of combined sildenafil and l-arginine during acute pulmonary embolism-induced pulmonary hypertension. Eur J Pharmacol 524:126–131PubMedCrossRefGoogle Scholar
  36. Tanus-Santos JE, Gordo WM, Cittadino M, Moreno H Jr (2000) Plasma cGMP levels in air embolism-induced acute lung injury. J Crit Care 15:137–141PubMedCrossRefGoogle Scholar
  37. Tanus-Santos JE, Desai M, Deak LR, Pezzullo JC, Abernethy DR, Flockhart DA, Freedman JE (2002) Effects of endothelial nitric oxide synthase gene polymorphisms on platelet function, nitric oxide release, and interactions with estradiol. Pharmacogenetics 12:407–413PubMedCrossRefGoogle Scholar
  38. Vaziri ND, Ding Y (2001) Effect of lead on nitric oxide synthase expression in coronary endothelial cells: role of superoxide. Hypertension 37:223–226PubMedGoogle Scholar
  39. Vaziri ND, Sica DA (2004) Lead-induced hypertension: role of oxidative stress. Curr Hypertens Rep 6:314–320PubMedGoogle Scholar
  40. Vaziri ND, Liang K, Ding Y (1999) Increased nitric oxide inactivation by reactive oxygen species in lead-induced hypertension. Kidney Int 56:1492–1498PubMedCrossRefGoogle Scholar
  41. Waldman SA, Murad F (1987) Cyclic GMP synthesis and function. Pharmacol Rev 39:163–196PubMedGoogle Scholar
  42. Wang XL, Mahaney MC, Sim AS, Wang J, Blangero J, Almasy L, Badenhop RB, Wilcken DE (1997) Genetic contribution of the endothelial constitutive nitric oxide synthase gene to plasma nitric oxide levels. Arterioscler Thromb Vasc Biol 17:3147–3153PubMedGoogle Scholar
  43. Yang BK, Vivas EX, Reiter CD, Gladwin MT (2003) Methodologies for the sensitive and specific measurement of S-nitrosothiols, iron-nitrosyls, and nitrite in biological samples. Free Radic Res 37:1–10PubMedCrossRefGoogle Scholar
  44. Zhou Y, Zanao RA, Barbosa F Jr, Parsons PJ, Krug FJ (2002) Investigations on a W-Rh permanent modifier for the detection of Pb in blood by electrothermal atomic absorption spectrometry. Spectrochim Acta Part B 57:1291–1300CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Fernando BarbosaJr
    • 1
  • Jonas T. C. Sertorio
    • 2
  • Raquel F. Gerlach
    • 3
  • Jose E. Tanus-Santos
    • 2
  1. 1.Department of Clinical, Toxicological and Food Science AnalysisFaculty of Pharmaceutical Sciences of Ribeirao PretoRibeirao PretoBrazil
  2. 2.Department of Pharmacology, Faculty of Medicine of Ribeirao PretoUniversity of Sao PauloRibeirao PretoBrazil
  3. 3.Department of Morphology, Estomatology and Physiology, Dental School of Ribeirao PretoUniversity of Sao Paulo Ribeirao PretoBrazil

Personalised recommendations