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Vol. 53, Issue 3, 483-491, March 1998

The Rat Heme Oxygenase-1 Gene Is Transcriptionally Induced via the Protein Kinase A Signaling Pathway in Rat Hepatocyte Cultures

Stephan Immenschuh,1 Thomas Kietzmann, Vera Hinke, Matthias Wiederhold, Norbert Katz, and Ursula Muller-Eberhard

Departments of Pediatrics, Biochemistry, and Pharmacology (S.I., U.M.-E.), Cornell University Medical College, New York, New York 10021, Institut für Klinische Chemie und Pathobiochemie (S.I., M.W., N.K.), Klinik der Justus-Liebig-Universität Giessen, D-35392 Giessen, Germany, and Institut für Biochemie und Molekulare Zellbiologie (T.K.), D-37073 Göttingen, Germany

Heme oxygenase-1 (HO-1) is the inducible form of the rate-limiting enzyme of heme degradation; it regulates the cellular content of heme. To investigate the role of the cAMP-dependent protein kinase (PKA) signaling pathway on hepatic HO-1 gene expression, primary rat hepatocyte cultures were treated with the PKA-stimulating agents dibutyryl-cAMP (Bt2cAMP), forskolin, and glucagon. HO-1 mRNA levels were induced by these agents in a time-dependent manner with a transient maximum after 6 hr of treatment. The induction of HO-1 was dose dependent, reaching a maximum at concentrations of 250 µM Bt2cAMP and 50 nM glucagon, respectively. The accumulation of HO-1 mRNA correlated with increased levels of HO-1 protein as determined by Western blot analysis. The Bt2cAMP-dependent induction of HO-1 mRNA expression was prevented by pretreatment with the PKA inhibitor KT5720 but not with the protein kinase G inhibitor KT5823. HO-1 mRNA induction by CdCl2 and heme was differentially affected by Bt2cAMP. Up-regulation of the HO-1 gene by Bt2cAMP occurred on the transcriptional level as determined by nuclear run-off assay and blocking of the Bt2cAMP-dependent induction of HO-1 mRNA by actinomycin D. Treatment with Bt2cAMP increased the half-life of HO-1 mRNA from 4.7 to 5.5 hr. Taken together, the results of the current study demonstrate that HO-1 gene expression is induced by activation of the cAMP signal transduction pathway via a transcriptional mechanism in primary rat hepatocyte cultures.

Copyright © 1998 by The American Society for Pharmacology and Experimental Therapeutics


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