Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

In vitro inhibition of heme oxygenase isoenzymes by metalloporphyrins

Abstract

Objective:

Neonatal jaundice results from an increased bilirubin production and decreased hepatic bilirubin conjugation and excretion. Severe hyperbilirubinemia is currently treated with phototherapy or exchange transfusion; however, its prevention by inhibiting bilirubin formation is a more logical strategy. Heme oxygenase (HO), with inducible (HO-1) and constitutive (HO-2) isoenzymes, is the rate-limiting enzyme in heme catabolism, producing equimolar amounts of bilirubin and carbon monoxide (CO). Metalloporphyrins (Mps) are heme derivatives that competitively inhibit HO and thereby suppress hyperbilirubinemia. No systematic studies have been reported evaluating whether the HO isoenzymes are inhibited differentially by various Mps. Identification of Mps that selectively inhibit the inducible HO-1 without affecting the ‘housekeeping’ HO-2 isoenzyme might be desirable in the clinical setting of hemolytic disease, in which the Hmox1 gene is greatly induced. Although bilirubin production is due to the activity of both HO-1 and HO-2, the inhibition of HO-1 with a relative sparing of HO-2 activity might provide the most selective approach for the treatment of hemolytic disease.

Study Design:

We determined for the deutero-, proto-, meso- and bis-glycol porphyrins with zinc, tin and chromium as central atoms, respectively, the concentration needed for 50% inhibition (I50) of HO-1 and HO-2 activities in rat spleen and brain tissue.

Result:

For a given Mp, HO-1 activity was less inhibited than that of HO-2. The order of inhibitor potency of each Mp was nearly identical for both isoenzymes. Tin mesoporphyrin was the most potent inhibitor for both isoenzymes. HO-2 selectivity was greatest for tin protoporphyrin. Conversely, the Zn compounds were least inhibitory toward HO-2. No Mp preferentially inhibited HO-1.

Conclusion:

Mps that produce a less inhibitory effect on HO-2, while limiting the response of the inducible HO-1, such as ZnPP, may be a useful clinical tool.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Vreman HJ, Mahoney JJ, Stevenson DK . Carbon monoxide and carboxyhemoglobin. Adv Pediatr 1995; 42: 303–325.

    CAS  Google Scholar 

  2. Coburn RF, Wallace HW, Abboud R . Redistribution of body carbon monoxide after hemorrhage. Amer J Physiol 1971; 220: 868–873.

    Article  CAS  Google Scholar 

  3. Landaw SA, Winchell HS, Boone RF . Measurement of endogenous carbon monoxide production in hemolytic disease of the inborn. Clin Res 1971; 19: 208A.

    Google Scholar 

  4. Valaes T, Drummond GS, Kappas A . Control of hyperbilirubinemia in glucose-6-phosphate dehydrogenase-deficient newborns using an inhibitor of bilirubin production, Sn mesoporphyrin. Pediatrics 1998; 101: E1.

    Article  CAS  Google Scholar 

  5. Lucey JF . Bilirubin and brain damage—a real mess. Pediatrics 1982; 69: 381–382.

    Google Scholar 

  6. Ennever JF . Phototherapy for neonatal jaundice. Yearly review. Photochem Photobiol 1988; 47: 871–876.

    Article  CAS  Google Scholar 

  7. Vreman HJ, Wong RJ, Stevenson DK, Route RK, Reader SD, Fejer MM et al. Light emitting diodes: a novel light source for phototherapy. Pediatr Res 1998; 44: 804–809.

    Article  CAS  Google Scholar 

  8. Lee KS, Gartner LM . Management of unconjugated hyperbilirubinemia in the newborn. Semin Liver Dis 1983; 3: 52–63.

    Article  CAS  Google Scholar 

  9. Mahoney JJ, Wong RJ, Vreman HJ, Stevenson DK . Fetal hemoglobin of transfused neonates and spectrophotometric measurements of oxyhemoglobin and carboxyhemoglobin. J Clin Monit 1991; 7: 154–160.

    Article  CAS  Google Scholar 

  10. Tenhunen R, Marver HS, Schmid R . The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc Natl Acad Sci USA 1968; 61: 748–755.

    Article  CAS  Google Scholar 

  11. Maines MD, Trakshel GM, Kutty RK . Characterization of two constitutive forms of rat liver microsomal heme oxygenase. Only one molecular species of the enzyme is inducible. J Biol Chem 1986; 261: 411–419.

    CAS  PubMed  Google Scholar 

  12. Cruse I, Maines MD . Evidence suggesting that the two forms of heme oxygenase are products of different genes. J Biol Chem 1988; 263: 3348–3353.

    CAS  PubMed  Google Scholar 

  13. Shibahara S, Yoshizawa M, Suzuki H, Takeda K, Meguro K, Endo K . Functional analysis of cDNAs for two types of human heme oxygenase and evidence for their separate regulation. J Biochem 1993; 113: 214–218.

    Article  CAS  Google Scholar 

  14. Vreman HJ, Wong RJ, Stevenson DK . Sources, sinks, and measurements of carbon monoxide. Carbon Monoxide and Cardiovascular Functions. CRC Press: Boca Raton, FL, 2001.

    Google Scholar 

  15. Raju VS, McCoubrey Jr WK, Maines MD . Regulation of heme oxygenase-2 by glucocorticoids in neonatal rat brain: characterization of a functional glucocorticoid response element. Biochim Biophys Acta 1997; 1351: 89–104.

    Article  CAS  Google Scholar 

  16. McCoubrey Jr WK, Huang TJ, Maines MD . Isolation and characterization of a cDNA from the rat brain that encodes hemoprotein heme oxygenase-3. Eur J Biochem 1997; 247: 725–732.

    Article  CAS  Google Scholar 

  17. Hintz SR, Kwong LK, Vreman HJ, Stevenson DK . Recovery of exogenous heme as carbon monoxide and biliary heme in adult rats after tin protoporphyrin treatment. J Pediatr Gastroenterol Nutr 1987; 6: 302–306.

    Article  CAS  Google Scholar 

  18. Vreman HJ, Ekstrand BC, Stevenson DK . Selection of metalloporphyrin heme oxygenase inhibitors based on potency and photoreactivity. Pediatr Res 1993; 33: 195–200.

    Article  CAS  Google Scholar 

  19. Qato MK, Maines MD . Prevention of neonatal hyperbilirubinaemia in non-human primates by Zn-protoporphyrin. Biochem J 1985; 226: 51–57.

    Article  CAS  Google Scholar 

  20. Vreman HJ, Lee OK, Stevenson DK . In vitro and in vivo characteristics of a heme oxygenase inhibitor: ZnBG. Am J Med Sci 1991; 302: 335–341.

    Article  CAS  Google Scholar 

  21. Kappas A, Simionatto CS, Drummond GS, Sassa S, Anderson KE . The liver excretes large amounts of heme into bile when heme oxygenase is inhibited competitively by Sn protoporphyrin. Proc Natl Acad Sci USA 1985; 82: 896–900.

    Article  CAS  Google Scholar 

  22. Vallier HA, Rodgers PA, Stevenson DK . Oral administration of zinc deuteroporphyrin IX 2,4 bis glycol inhibits heme oxygenase in neonatal rats. Dev Pharmacol Ther 1991; 17: 220–222.

    Article  CAS  Google Scholar 

  23. Vallier HA, Rodgers PA, Stevenson DK . Inhibition of heme oxygenase after oral vs intraperitoneal administration of chromium porphyrins. Life Sci 1993; 52: L79–L84.

    Article  Google Scholar 

  24. Vreman HJ, Cipkala DA, Stevenson DK . Characterization of porphyrin heme oxygenase inhibitors. Can J Physiol Pharmacol 1996; 74: 278–285.

    CAS  PubMed  Google Scholar 

  25. Marks GS, Brien JF, Nakatsu K, McLaughlin BE . Does carbon monoxide have a physiological function? Trends Pharmacol Sci 1991; 12: 185–188.

    Article  CAS  Google Scholar 

  26. Stocker R, Yamamoto Y, McDonagh AF, Glazer AN, Ames BN . Bilirubin is an antioxidant of possible physiological importance. Science 1987; 235: 1043–1046.

    Article  CAS  Google Scholar 

  27. Dennery PA, McDonagh AF, Spitz DR, Rodgers PA, Stevenson DK . Hyperbilirubinemia results in reduced oxidative injury in neonatal Gunn rats exposed to hyperoxia. Free Radic Biol Med 1995; 19: 395–404.

    Article  CAS  Google Scholar 

  28. Vreman HJ, Stevenson DK . Heme oxygenase activity as measured by carbon monoxide production. Anal Biochem 1988; 168: 31–38.

    Article  CAS  Google Scholar 

  29. Vreman HJ, Stevenson DK . Detection of heme oxygenase activity by measurement of CO. In: Maines MD, Costa LG, Reed DJ, Sassa S, Sipes IG (eds). Current Protocols in Toxicology. John Wiley & Sons, Inc.: New York, 1999, pp 9.2.1–9.2.10.

    Google Scholar 

  30. Vreman HJ, Wong RJ, Kim EC, Nabseth DC, Marks GS, Stevenson DK . Haem oxygenase activity in human umbilical cord and rat vascular tissues. Placenta 2000; 21: 337–344.

    Article  CAS  Google Scholar 

  31. McCoubrey Jr WK, Huang TJ, Maines MD . Heme oxygenase-2 is a hemoprotein and binds heme through heme regulatory motifs that are not involved in heme catalysis. J Biol Chem 1997; 272: 12568–12574.

    Article  CAS  Google Scholar 

  32. Kinobe RT, Vlahakis JZ, Vreman HJ, Stevenson DK, Brien JF, Szarek WA et al. Selectivity of imidazole-dioxolane compounds for in vitro inhibition of microsomal haem oxygenase isoforms. Br J Pharmacol 2006; 147: 307–315.

    Article  CAS  Google Scholar 

  33. Vlahakis JZ, Kinobe RT, Bowers RJ, Brien JF, Nakatsu K, Szarek WA . Imidazoledioxolane compounds as isozyme-selective heme oxygenase inhibitors. J Med Chem 2006; 49: 4437–4441.

    Article  CAS  Google Scholar 

  34. Morisawa T, Wong RJ, Bhutani VK, Vreman HJ, Stevenson DK . Inhibition of heme oxygenase activity in newborn mice by azalanstat. Can J Physiol Pharmacol 2008; 86: 651–659.

    Article  CAS  Google Scholar 

  35. Zhang Y, Furuyama K, Adachi T, Ishikawa K, Matsumoto H, Masuda T et al. Hypoxemia and attenuated hypoxic ventilatory responses in mice lacking heme oxygenase-2: evidence for a novel role of heme oxygenase-2 as an oxygen sensor. Adv Exp Med Biol 2006; 580: 161–166; discussion 351–169.

    Article  CAS  Google Scholar 

  36. Kemp PJ . Hemeoxygenase-2 as an O2 sensor in K+ channel-dependent chemotransduction. Biochem Biophys Res Commun 2005; 338: 648–652.

    Article  CAS  Google Scholar 

  37. Vreman HJ, Wong RJ, Stevenson DK . Alternative metalloporphyrins for the treatment of neonatal jaundice. J Perinatol 2001; 21 (Suppl 1): S108–S113.

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported in part by the National Institutes of Health (Grants HD 14426, HD/HL 58013, HL 68703), the Hess Research Fund, the Mary L Johnson Research Fund and the HM Lui Research Fund.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R J Wong.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wong, R., Vreman, H., Schulz, S. et al. In vitro inhibition of heme oxygenase isoenzymes by metalloporphyrins. J Perinatol 31 (Suppl 1), S35–S41 (2011). https://doi.org/10.1038/jp.2010.173

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/jp.2010.173

Keywords

This article is cited by

Search

Quick links