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Research ArticleINFLAMMATION AND IMMUNOPHARMACOLOGY

Adenoviral Vector-Mediated Transfer of Human Heme Oxygenase in Rats Decreases Renal Heme-Dependent Arachidonic Acid Epoxygenase Activity

Nader G. Abraham, Shanlong Jiang, Liming Yang, Barbara A. Zand, Michal Laniado-Schwartzman, Jackleen Marji, George S. Drummond and Attallah Kappas
Journal of Pharmacology and Experimental Therapeutics May 2000, 293 (2) 494-500;
Nader G. Abraham
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Shanlong Jiang
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Liming Yang
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Barbara A. Zand
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Michal Laniado-Schwartzman
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Jackleen Marji
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George S. Drummond
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Attallah Kappas
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Abstract

Intravenous administration of an adenovirus human heme oxygenase (HO)-1 gene construct to rats resulted in functional expression of human HO-1 in brain, heart, lung, liver, and kidney. Because accurate assessment of human HO-1 mRNA in various tissues by Northern analysis is not sufficiently sensitive, we developed a method for quantifying human HO-1 mRNA copies with quantitative reverse transcription- polymerase chain reaction techniques; this allowed us to use the same primers for both the sample and internal standard. Administration of the adenovirus human HO-1 gene resulted in the detection of human HO-1 mRNA in various tissues with the highest levels seen in the kidney followed, in order, by lung > liver > brain > heart. Human HO-1 was detectable for up to 4 weeks in all tissues studied. Administration of adenovirus human HO-1 resulted in maximal increase of HO activity after 1 to 2 weeks in rats. The increase in HO activity due to gene transfer also was associated with a parallel decrease (∼25%) in cytochrome P-450 (CYP) content and in CYP-dependant arachidonic acid metabolism. In addition, we investigated the possibility that the human HO-1 gene altered the expression of the endogenous rat enzyme after administration of cobalt chloride s.c.. Cobalt chloride administration resulted in increased HO activity in all tissues examined in rats transduced with the human HO-1 gene to the same degree as in nontransduced rats. The metal was a more potent inducer of renal HO activity than was the adenoviral-mediated human HO-1 vector. The increase in HO activity after adenoviral-mediated human HO-1 transfer was associated with a decrease in microsomal heme-CYP and CYP activity. The increase in HO-1 activity after adenovirus-mediated human HO-1 gene transfer may prove useful as a means of selectively increasing enzyme activity in a specific organ and regulating homeostasis by modulation of vasoactive molecules such as carbon monoxide and bilirubin and, in addition, providing a means of delivering the human HO-1 gene for experimental purposes.

Footnotes

  • Send reprint requests to: Nader G. Abraham, Department of Pharmacology, New York Medical College, Valhalla, NY 10595. E-mail: nader_abraham{at}nymc.edu

  • ↵1 This study was supported in part by Grants RO1 HL54138 and EY06531 from the National Institutes of Health.

  • Abbreviations:
    CYP
    cytochrome P-450
    HO
    heme oxygenase
    PCR
    polymerase chain reaction
    pfu
    plaque-forming unit
    RT
    reverse transcription
    bp
    base pair(s)
    • Received September 14, 1999.
    • Accepted January 17, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 293 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 293, Issue 2
1 May 2000
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Research ArticleINFLAMMATION AND IMMUNOPHARMACOLOGY

Adenoviral Vector-Mediated Transfer of Human Heme Oxygenase in Rats Decreases Renal Heme-Dependent Arachidonic Acid Epoxygenase Activity

Nader G. Abraham, Shanlong Jiang, Liming Yang, Barbara A. Zand, Michal Laniado-Schwartzman, Jackleen Marji, George S. Drummond and Attallah Kappas
Journal of Pharmacology and Experimental Therapeutics May 1, 2000, 293 (2) 494-500;

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Research ArticleINFLAMMATION AND IMMUNOPHARMACOLOGY

Adenoviral Vector-Mediated Transfer of Human Heme Oxygenase in Rats Decreases Renal Heme-Dependent Arachidonic Acid Epoxygenase Activity

Nader G. Abraham, Shanlong Jiang, Liming Yang, Barbara A. Zand, Michal Laniado-Schwartzman, Jackleen Marji, George S. Drummond and Attallah Kappas
Journal of Pharmacology and Experimental Therapeutics May 1, 2000, 293 (2) 494-500;
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