Heme oxygenase isoform-specific expression and distribution in the rat kidney

Jean Louis Da Silva, Barbara A. Zand, Li Ming Yang, Hatem Sabaawy, Elias Lianos, Nader G. Abraham

Research output: Contribution to journalArticle

88 Citations (Scopus)

Abstract

Background. The heme oxygenase (HO) genes, HO-1 and HO-2, are the limiting steps in heme degradation and in the regulation of renal heme-dependent enzymes. Previously we reported that selective overexpression of renal HO-1 resulted in a decrease of microsomal heme and the cytochrome P450-dependent arachidonic acid metabolite, 20 HETE, a vasoconstrictor. The present study was undertaken to explore the relative expression and contribution of each of the HO isoforms to HO activity in the rat kidney. Methods and Results. Renal HO activity increased above control levels after an injection of the inducers of HO activity, heme or SnCl 2 . Stannous Mesoporphyrin (SnMP), a nonselective inhibitor of HO, when used alone or in combination with heme or SnCl 2 , decreased HO activity. Heme alone and combined with SnCl 2 decreased the levels of heme content by 13 and 35%, respectively. Western blot analysis showed that both SnCl 2 and heme readily induced HO-1 protein, whereas HO-2 was constitutively expressed. Immunohistochemistry showed the distribution of the HO-1 isoform primarily in proximal convoluted tubules. Western blot analysis exhibited relatively higher levels of HO-1 in isolated proximal tubules and relatively higher HO-2 levels in the thick ascending limbs of the loop of Henle and preglomerular arterioles. In vivo administration of HO-1 and HO-2 antisense oligodeoxynucleotides further confirmed that HO-2, but not HO-1, contributed to the basal HO activity; however, following induction of HO with heme, antisense to HO-1, but not to HO-2, inhibited the induced levels of HO activity. Conclusion. These results suggest that HO-2 is constitutively expressed in the rat kidney mainly within tubular and arteriolar structures, and its activity may modulate physiological function under basal conditions. On the other hand, the basal levels of expression of HO-1 in the rat kidney are relatively low, and its contribution to HO activity and the regulation of hemoproteins such as cytochrome P450 become apparent only under pathophysiological conditions causing HO induction.

Original languageEnglish (US)
Pages (from-to)1448-1457
Number of pages10
JournalKidney International
Volume59
Issue number4
DOIs
StatePublished - Jan 1 2001

Fingerprint

Heme Oxygenase (Decyclizing)
Heme Oxygenase-1
Protein Isoforms
Heme
Kidney
Cytochrome P-450 Enzyme System
Western Blotting
Loop of Henle
Oligodeoxyribonucleotides
Arterioles
Vasoconstrictor Agents
heme oxygenase-2
Arachidonic Acid

All Science Journal Classification (ASJC) codes

  • Nephrology

Cite this

Da Silva, Jean Louis ; Zand, Barbara A. ; Yang, Li Ming ; Sabaawy, Hatem ; Lianos, Elias ; Abraham, Nader G. / Heme oxygenase isoform-specific expression and distribution in the rat kidney. In: Kidney International. 2001 ; Vol. 59, No. 4. pp. 1448-1457.
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abstract = "Background. The heme oxygenase (HO) genes, HO-1 and HO-2, are the limiting steps in heme degradation and in the regulation of renal heme-dependent enzymes. Previously we reported that selective overexpression of renal HO-1 resulted in a decrease of microsomal heme and the cytochrome P450-dependent arachidonic acid metabolite, 20 HETE, a vasoconstrictor. The present study was undertaken to explore the relative expression and contribution of each of the HO isoforms to HO activity in the rat kidney. Methods and Results. Renal HO activity increased above control levels after an injection of the inducers of HO activity, heme or SnCl 2 . Stannous Mesoporphyrin (SnMP), a nonselective inhibitor of HO, when used alone or in combination with heme or SnCl 2 , decreased HO activity. Heme alone and combined with SnCl 2 decreased the levels of heme content by 13 and 35{\%}, respectively. Western blot analysis showed that both SnCl 2 and heme readily induced HO-1 protein, whereas HO-2 was constitutively expressed. Immunohistochemistry showed the distribution of the HO-1 isoform primarily in proximal convoluted tubules. Western blot analysis exhibited relatively higher levels of HO-1 in isolated proximal tubules and relatively higher HO-2 levels in the thick ascending limbs of the loop of Henle and preglomerular arterioles. In vivo administration of HO-1 and HO-2 antisense oligodeoxynucleotides further confirmed that HO-2, but not HO-1, contributed to the basal HO activity; however, following induction of HO with heme, antisense to HO-1, but not to HO-2, inhibited the induced levels of HO activity. Conclusion. These results suggest that HO-2 is constitutively expressed in the rat kidney mainly within tubular and arteriolar structures, and its activity may modulate physiological function under basal conditions. On the other hand, the basal levels of expression of HO-1 in the rat kidney are relatively low, and its contribution to HO activity and the regulation of hemoproteins such as cytochrome P450 become apparent only under pathophysiological conditions causing HO induction.",
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Heme oxygenase isoform-specific expression and distribution in the rat kidney. / Da Silva, Jean Louis; Zand, Barbara A.; Yang, Li Ming; Sabaawy, Hatem; Lianos, Elias; Abraham, Nader G.

In: Kidney International, Vol. 59, No. 4, 01.01.2001, p. 1448-1457.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Heme oxygenase isoform-specific expression and distribution in the rat kidney

AU - Da Silva, Jean Louis

AU - Zand, Barbara A.

AU - Yang, Li Ming

AU - Sabaawy, Hatem

AU - Lianos, Elias

AU - Abraham, Nader G.

PY - 2001/1/1

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N2 - Background. The heme oxygenase (HO) genes, HO-1 and HO-2, are the limiting steps in heme degradation and in the regulation of renal heme-dependent enzymes. Previously we reported that selective overexpression of renal HO-1 resulted in a decrease of microsomal heme and the cytochrome P450-dependent arachidonic acid metabolite, 20 HETE, a vasoconstrictor. The present study was undertaken to explore the relative expression and contribution of each of the HO isoforms to HO activity in the rat kidney. Methods and Results. Renal HO activity increased above control levels after an injection of the inducers of HO activity, heme or SnCl 2 . Stannous Mesoporphyrin (SnMP), a nonselective inhibitor of HO, when used alone or in combination with heme or SnCl 2 , decreased HO activity. Heme alone and combined with SnCl 2 decreased the levels of heme content by 13 and 35%, respectively. Western blot analysis showed that both SnCl 2 and heme readily induced HO-1 protein, whereas HO-2 was constitutively expressed. Immunohistochemistry showed the distribution of the HO-1 isoform primarily in proximal convoluted tubules. Western blot analysis exhibited relatively higher levels of HO-1 in isolated proximal tubules and relatively higher HO-2 levels in the thick ascending limbs of the loop of Henle and preglomerular arterioles. In vivo administration of HO-1 and HO-2 antisense oligodeoxynucleotides further confirmed that HO-2, but not HO-1, contributed to the basal HO activity; however, following induction of HO with heme, antisense to HO-1, but not to HO-2, inhibited the induced levels of HO activity. Conclusion. These results suggest that HO-2 is constitutively expressed in the rat kidney mainly within tubular and arteriolar structures, and its activity may modulate physiological function under basal conditions. On the other hand, the basal levels of expression of HO-1 in the rat kidney are relatively low, and its contribution to HO activity and the regulation of hemoproteins such as cytochrome P450 become apparent only under pathophysiological conditions causing HO induction.

AB - Background. The heme oxygenase (HO) genes, HO-1 and HO-2, are the limiting steps in heme degradation and in the regulation of renal heme-dependent enzymes. Previously we reported that selective overexpression of renal HO-1 resulted in a decrease of microsomal heme and the cytochrome P450-dependent arachidonic acid metabolite, 20 HETE, a vasoconstrictor. The present study was undertaken to explore the relative expression and contribution of each of the HO isoforms to HO activity in the rat kidney. Methods and Results. Renal HO activity increased above control levels after an injection of the inducers of HO activity, heme or SnCl 2 . Stannous Mesoporphyrin (SnMP), a nonselective inhibitor of HO, when used alone or in combination with heme or SnCl 2 , decreased HO activity. Heme alone and combined with SnCl 2 decreased the levels of heme content by 13 and 35%, respectively. Western blot analysis showed that both SnCl 2 and heme readily induced HO-1 protein, whereas HO-2 was constitutively expressed. Immunohistochemistry showed the distribution of the HO-1 isoform primarily in proximal convoluted tubules. Western blot analysis exhibited relatively higher levels of HO-1 in isolated proximal tubules and relatively higher HO-2 levels in the thick ascending limbs of the loop of Henle and preglomerular arterioles. In vivo administration of HO-1 and HO-2 antisense oligodeoxynucleotides further confirmed that HO-2, but not HO-1, contributed to the basal HO activity; however, following induction of HO with heme, antisense to HO-1, but not to HO-2, inhibited the induced levels of HO activity. Conclusion. These results suggest that HO-2 is constitutively expressed in the rat kidney mainly within tubular and arteriolar structures, and its activity may modulate physiological function under basal conditions. On the other hand, the basal levels of expression of HO-1 in the rat kidney are relatively low, and its contribution to HO activity and the regulation of hemoproteins such as cytochrome P450 become apparent only under pathophysiological conditions causing HO induction.

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