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Journal of Histochemistry and Cytochemistry, Vol. 46, 1151-1160, October 1998, Copyright © 1998, The Histochemical Society, Inc.

ARTICLE

Effect of 3-Methylcholanthrene Administration on Expression of Cytochrome P-450 Isoforms Induced by Phenobarbital in Rat Hepatocytes

Kazuto Minoa, Jun Watanabea, and Shinsuke Kanamuraa
a Department of Anatomy, Kansai Medical University, Moriguchi, Osaka, Japan

Correspondence to: Shinsuke Kanamura, Dept. of Anatomy, Kansai Medical Univ., 10-15 Fumizono-cho, Moriguchi, Osaka 570, Japan..

The effects of an inducer on expression of cytochrome P-450 (P-450) isoforms induced antecedently by another inducer are unknown. Thus, we examined the amount of phenobarbital (PB)-inducible P-450 isoforms (P-450 2B1/2B2) in hepatocytes from rats injected first with PB and then with 3-methylcholanthrene (MC) (PB+MC-treated animals) by quantitative immunohistochemistry. In addition, expression of P-450 2B2 mRNA was examined by in situ hybridization. In PB-treated animals, P-450 2B1/2B2 content increased in perivenular and midzonal hepatocytes. In PB+MC-treated animals, however, the PB-induced increase in 2B1/2B2 content was suppressed in perivenular hepatocytes but promoted in midzonal hepatocytes. The hybridization signal for P-450 2B2 mRNA appeared almost exclusively in perivenular hepatocytes after 24 hr of PB injection and disappeared after 48 hr of injection. In PB+MC-treated animals, however, strong hybridization signal was observed in midzonal and perivenular hepatocytes after 48 hr of PB injection. The promotion of the increase in P-450 2B1/2B2 content in midzonal hepatocytes in PB+MC-treated animals probably corresponds to the strong hybridization signal, whereas there appeared to be a divergence between the intensity of the signal and the content in perivenular hepatocytes. The results indicate that MC administration drastically influences the pattern of expression of P-450 isoforms induced by PB in perivenular and midzonal hepatocytes. (J Histochem Cytochem 46:1151–1160, 1998)

Key Words: cytochrome P-450 2B, phenobarbital, 3-methylcholanthrene, quantitative immunohistochemistry, in situ hybridization, liver, rat


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