Abstract
Zinc protoporphyrin IX (ZnPP) has been shown to inhibit heme oxygenase (HO) activity effectively in vivo and has potential in the treatment of neonatal jaundice. Because this is a transitional or temporary condition lasting only several days, an effective chemopreventive agent with a relatively short duration of action would be desirable for the treatment of severe neonatal jaundice. To determine the effective duration of action of ZnPP, we administered either 40 nmol/g of body weight ZnPP or 5 μL/g body weight diluent intraperitoneally to neonatal rats 24-36 h after birth. Between 0 and 21 d after ZnPP dosing, the duration of action was investigated through measurements of serum bilirubin and hepatic and splenic HO inhibition, which were correlated to measurements of ZnPP distribution. Significant (p< 0.05) hepatic HO inhibition, ranging from 27 to 51%, was observed in the liver between 1 and 4 d after dosing, concurrent with a 23-28% reduction in serum bilirubin levels, and was associated with ZnPP tissue concentrations of 27-38 nmol/g. Splenic HO was not inhibited measurably by the much lower concentrations of ZnPP found in the spleen (2.8-20.1 nmol/g) between 0 and 21 d after dosing. Furthermore, HO isoform 1 (HO-1) induction was apparently not a confounding factor in the duration of action of ZnPP, because the modest increases in HO-1 protein levels were not sustained longer than 24 h after ZnPP administration. Our findings demonstrated that the duration of action of ZnPP in neonatal rats is less than 1 wk. The reduction in serum bilirubin levels, the short duration of action and minimal confounding effects suggest that ZnPP may be an effective chemopreventive agent for the treatment of severe neonatal jaundice.
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Abbreviations
- HO:
-
heme oxygenase
- HO-1 and -2:
-
heme oxygenase isoforms 1 and 2
- ZnPP:
-
zinc protoporphyrin
- SnPP:
-
tin protoporphyrin
- SnMP:
-
tin mesoporphyrin
- GAPD:
-
glyceraldehyde-3-phosphate dehydrogenase
- B.W.:
-
body weight
- i.p.:
-
intraperitoneal
References
Maisels MJ 1988 Neonatal jaundice. Semin Liver Dis 8: 148–162.
Ostrow JD 1988 Therapeutic amelioration of jaundice: old and new strategies. Hepatology 8: 683–689.
Tenhunen R, Marver HS, Schmid R 1968 The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc Natl Acad Sci USA 61: 748–755.
Shibahara S, Yoshizawa M, Suzuki H, Takeda K, Meguro K, Endo K 1993 Functional analysis of cDNAs for two types of human heme oxygenase and evidence for their separate regulation. J Biochem 113: 214–218.
Maines MD, Trakshel GM, Kutty RK 1986 Characterization of two constitutive forms of rat liver microsomal heme oxygenase. Only one molecular species of the enzyme is inducible. J Biol Chem 261: 411–419.
Drummond GS, Kappas A 1981 Prevention of neonatal hyperbilirubinemia by tin protoporphyrin IX, a potent competitive inhibitor of heme oxidation. Proc Natl Acad Sci USA 78: 6466–6470.
Drummond GS, Galbraith RA, Sardana MK, Kappas A 1987 Reduction of the C2 and C4 vinyl groups of Sn-protoporphyrin to form Sn-mesoporphyrin markedly enhances the ability of the metalloporphyrin to inhibit in vivo heme catabolism. Arch Biochem Biophys 255: 64–74.
Milleville GS, Levitt MD, Engel RR 1985 Tin protoporphyrin inhibits carbon monoxide production in adult mice. Pediatr Res 19: 94–96.
Kappas A, Drummond GS, Manola T, Petmezaki S, Valaes T 1988 Sn-protoporphyrin use in the management of hyperbilirubinemia in term newborns with direct Coombs-positive ABO incompatibility. Pediatrics 81: 485–497.
Valaes T, Petmezaki S, Henschke C, Drummond GS, Kappas A 1994 Control of jaundice in preterm newborns by an inhibitor of bilirubin production: studies with tin-mesoporphyrin. Pediatrics 93: 1–11.
Rubaltelli FF, Guerrini P, Reddi E, Jori G 1989 Tin-protoporphyrin in the management of children with Crigler-Najjar disease.[See comments. ] Pediatrics 84: 728–731.
Galbraith RA, Drummond GS, Kappas A 1992 Suppression of bilirubin production in the Crigler-Najjar type I syndrome: studies with the heme oxygenase inhibitor tin-mesoporphyrin. [See comments. ] Pediatrics 89: 175–182.
Galbraith RA, Kappas A 1989 Pharmacokinetics of tin-mesoporphyrin in man and the effects of tin-chelated porphyrins on hyperexcretion of heme pathway precursors in patients with acute inducible porphyria. Hepatology 9: 882–888.
Kappas A, Drummond GS, Galbraith RA 1993 Prolonged clinical use of a heme oxygenase inhibitor: hematological evidence for an inducible but reversible irondeficiency state. Pediatrics 91: 537–539.
Fort FL, Gold J 1989 Phototoxicity of tin protoporphyrin, tin mesoporphyrin, and tin diiododeuteroporphyrin under neonatal phototherapy conditions. Pediatrics 84: 1031–1037.
Hintz SR, Vreman HJ, Stevenson DK 1990 Mortality of metalloporphyrin-treated neonatal rats after light exposure. Dev Pharmacol Ther 14: 187–192.
Drummond GS, Rosenberg DW, Kihlstrom JA, Kappas A 1989 Effects of tinporphyrins on developmental changes in hepatic cytochrome P450 content, selected P450-dependent drug-metabolizing enzyme activities and brain glutathione levels in the newborn rat. Pharmacology 39: 273–284.
Drummond GS, Kappas A 1986 Sn-protoporphyrin inhibition of fetal and neonatal brain heme oxygenase. Transplacental passage of the metalloporphyrin and prenatal suppression of hyperbilirubinemia in the newborn animal. J Clin Invest 77: 971–976.
Mark JA, Maines MD 1992 Tin-protoporphyrin-mediated disruption in vivo of heme oxygenase-2 protein integrity and activity in rat brain. Pediatr Res 32: 324–329.
Maines MD, Trakshel GM 1992 Differential regulation of heme oxygenase isozymes by Sn- and Zn-protoporphyrins: possible relevance to suppression of hyperbilirubinemia. Biochim Biophys Acta 1131: 166–174.
Maines MD, Trakshel GM 1992 Tin-protoporphyrin: a potent inhibitor of hemoprotein-dependent steroidogenesis in rat adrenals and testes. J Pharmacol Exp Ther 260: 909–916.
Stout DL, Becker FF 1988 The effects of tin-protoporphyrin administration on hepatic xenobiotic metabolizing enzymes in the juvenile rat. Drug Metab Dispos 16: 23–26.
Sardana MK, Kappas A 1987 Dual control mechanism for heme oxygenase: tin(IV)-protoporphyrin potently inhibits enzyme activity while markedly increasing content of enzyme protein in liver. Proc Natl Acad Sci USA 84: 2464–2468.
Vreman HJ, Ekstrand BC, Stevenson DK 1993 Selection of metalloporphyrin heme oxygenase inhibitors based on potency and photoreactivity. Pediatr Res 33: 195–200.
Vreman HJ, Lee OK, Stevenson DK 1991 In vitro and in vivo characteristics of a heme oxygenase inhibitor: ZnBG. Am J Med Sci 302: 335–341.
Maines MD 1981 Zinc protoporphyrin is a selective inhibitor of heme oxygenase activity in the neonatal rat. Biochim Biophys Acta 673: 339–350.
Russo SM, Pepe JA, Donohue S, Cable EE, Lambrecht RW, Bonkovsky HL 1995 Tissue distribution of zinc-mesoporphyrin in rats: relationship to inhibition of heme oxygenase. J Pharmacol Exp Ther 272: 766–774.
Drummond GS, Kappas A 1982 Suppression of hyperbilirubinemia in the rat neonate by chromium-protoporphyrin. Interactions of metalloporphyrins with microsomal heme oxygenase of human spleen. J Exp Med 156: 1878–1883.
Rodgers PA, Vreman HJ, Stevenson DK 1990 Heme catabolism in rhesus neonates inhibited by zinc protoporphyrin. Dev Pharmacol Ther 14: 216–222.
Hamori CJ, Vreman HJ, Stevenson DK 1988 Suppression of carbon monoxide excretion by zinc mesoporphyrin in adult Wistar rats: evidence for potent in vivo inhibition of bilirubin production. Res Commun Chem Pathol Pharmacol 62: 41–48.
Qato MK, Maines MD 1985 Prevention of neonatal hyperbilirubinaemia in non-human primates by Zn-protoporphyrin. Biochem J 226: 51–57.
Vreman HJ, Gillman MJ, Downum KR, Stevenson DK 1990 In vitro generation of carbon monoxide from organic molecules and synthetic metalloporphyrins mediated by light. Dev Pharmacol Ther 15: 112–124.
Lutton J, Chertkov J, Levere R, Abraham N 1991 Comparative effect of heme analogues on hematopoiesis in lymphoproliferative disorders. Leuk Lymphoma 5: 179–185.
Ny L, Andersson KE, Grundemar L 1995 Inhibition by zinc protoporphyrin-IX of receptor-mediated relaxation of the rat aorta in a manner distinct from inhibition of haem oxygenase. Br J Pharmacol 115: 186–190.
Maines MD 1992 Heme Oxygenase: Clinical Applications and Functions. CRC Press, Boca Raton, FL, 248–249.
Vallier HA, Rodgers PA, Castillo RO, Stevenson DK 1991 Absorption of zinc deuteroporphyrin IX 2:4-bis-glycol by the neonatal rat small intestine in vivo. Dev Pharmacol Ther 17: 109–115.
Chomczynski P, Sacchi N 1987 Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159.
Vreman HJ, Stevenson DK 1988 Heme oxygenase activity as measured by carbon monoxide production. Anal Biochem 168: 31–38.
Lowry O, Rosebrough H, Farr A, Randall R 1951 Protein measurement with the Folin phenol reagent. J Biol Chem 193: 256–272.
Nakamura H, Lee Y 1977 Microdetermination of unbound bilirubin in icteric newborn sera: an enzymatic method employing peroxidase and glucose oxidase. Clin Chim Acta 79: 411–417.
Schwartz S, Stephenson B, Sarkar D, Freyholtz H, Ruth G 1980 Quantitative assay of erythrocyte “free” and zinc-protoporphyrin: clinical and genetic studies. Int J Biochem 12: 1053–1057.
Shibahara S, Muller R, Taguchi H, Yoshida T 1985 Cloning and expression of cDNA for rat heme oxygenase. Proc Natl Acad Sci USA 82: 7865–7869.
Adams M, Dubnick M, Kerlavage A, Moreno R, Kelley J, Utterback T, Nagle J 1992 Sequence identification of 2:375 human brain genes. Nature 355: 632–634.
Sambrook J, Fritsch E, Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Feinberg AP, Vogelstein B 1984 A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–272.
Laemmli UK 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Towbin H, Staehelin T, Gordon J 1979 Electrophoretic transfer of protein from polyacrylamide gels to nitrocelluose sheets:procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354.
Wilks A, Ortiz, DE, Montellano, PR 1993 Rat liver heme oxygenase. High level expression of a truncated soluble form and nature of the meso-hydroxylating species. J Biol Chem 268. 22357–22362
Maines MD, Kappas A 1975 Study of the developmental pattern of heme catabolism in liver and the effects of cobalt on cytochrome P-450 and the rate of heme oxidation during the neonatal period. J Exp Med 141: 1400–1410.
Hamori CJ, Vreman HJ, Rodgers PA, Stevenson DK 1989 Zinc protoporphyrin inhibits CO production in rats. J Pediatr Gastroenterol Nutr 8: 110–115.
Vreman HJ, Rodgers PA, Stevenson DK 1990 Zinc protoporphyrin administration for suppression of increased bilirubin production by iatrogenic hemolysis in rhesus neonates. J Pediatr 292: 7
Anderson KE, Simionatto CS, Drummond GS, Kappas A 1984 Tissue distribution and disposition of tin-protoporphyrin, a potent competitive inhibitor of heme oxygenase. J Pharmacol Exp Ther 228: 327–333.
Verma A, Hirsch DJ, Glatt CE, Ronnett GV, Snyder SH 1993 Carbon monoxide: a putative neural messenger [see comments]. Science 259: 381–384.
Zhuo M, Small S, Kandel E, Hawkins R 1993 Nitric oxide and carbon monoxide produce activity-dependent long-term synaptic enhancement in hippocampus. Science 260: 1946–1950.
Lin JH, Villalon P, Nelson JC, Abraham NG 1989 Expression of rat liver heme oxygenase gene during development. Arch Biochem Biophys 270: 623–629.
Galbraith R, Kappas A 1989 Regulation of food intake and body weight by cobalt porphyrins in animals. Proc Natl Acad Sci USA 86: 7653–7657.
Acknowledgements
The authors gratefully acknowledge the outstanding technical assistance of Christen Lee, Allie Shon, Ido Paz, and Vida Shokoohi.
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Supported by the National Institutes of Health Grant HD14426 and Mary L. Johnson Research Fund.
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Rodgers, P., Seidman, D., Wei, P. et al. Duration of Action and Tissue Distribution of Zinc Protoporphyrin in Neonatal Rats. Pediatr Res 39, 1041–1049 (1996). https://doi.org/10.1203/00006450-199606000-00018
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DOI: https://doi.org/10.1203/00006450-199606000-00018
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