Interactions of melatonin with the liver microsomal cytochrome P450 system of rats and humans in vitro and effects on the P450 system and the antioxidative status in rat liver after acute treatment

  • W. Klinger
  • E. Karge
  • U. Demme
  • M. Kretzschmar
Article
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Summary

In vitro melatonin binds to human and rat liver microsomal cytochrome P-450 (P450) according to a type II substrate. The affinity is similar to that of aniline with a general left-shift. Melatonin interferes with model monooxygenase reactions indicative of different P450 forms in humans and rats (in humans according to the lower specific P450 content less pronounced): the strongest inhibition was found for ethoxyresorufin O-deethylation, indicating the binding to P450 1A, the binding to P450 2B (ethoxycoumarin O-deethylation) was less pronounced, the least inhibition was found for P450 3A (ethylmorphine N-demethylation) reaction. The oxidase function was also inhibited: luminol amplified chemiluminescence was more inhibited than the lucigenin amplified one, hydrogen peroxide formation was inhibited at concentrations higher than 10−4 M, microsomal NADPH/Fe stimulated lipid peroxidation was inhibited at concentrations higher than 105 M.

In vivo melatonin prolonged hexobarbital sleeping time in rats in a dose dependent manner (ip. co-administration of 1, 5 and 20 mg/kg b.w. melatonin with 100 mg/kg hexobarbital). Immediately after awakening the animals were sacrificed: a small increase in P450 concentrations cannot be explained, no changes in P450 monooxygenase or oxidase activities nor in microsomal lipid peroxidation or GSH status could be observed.

Keywords

Melatonin P450 monooxygenase oxidase reactive oxygen species chemiluminescence lipid peroxidation GSH hexobarbital rat 

Abbreviations

CL

chemiluminescence

CPM

counts per minute

ECOD

ethoxycoumarin O-deethylation

EMND

ethylmorphine N-demethylation

EROD

ethoxyresorufin O-deethylation

GSH

glutathione, reduced

GSSG

glutathione disulfide, oxidized glutathione

H2O2

hydrogen peroxide

HB

hexobarbital (-Na)

HBST

hexobarbital sleeping time

LC

lucigenin

LM

luminol

LPO

lipid peroxidation

P450

cytochrome P-450

RLU

relative light unit

ROS

reactive oxygen species

TBARS

thiobarbituric acid reactive substances

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Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  • W. Klinger
    • 1
  • E. Karge
    • 1
  • U. Demme
    • 2
  • M. Kretzschmar
    • 3
  1. 1.Institute of Pharmacology and ToxicologyFriedrich Schiller University JenaJenaGermany
  2. 2.Institute of Forensic MedicineFriedrich Schiller University JenaJenaGermany
  3. 3.Clinic of Anaesthesiology and Intensive MedicineTown Hospital GeraJenaGermany

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