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Journal of Lipid Research, Vol. 44, 859-866, April 2003
Copyright © 2003 by Lipid Research, Inc.
Methods |
Monitoring hepatic cholesterol 7
-hydroxylase activity by assay of the stable bile acid intermediate 7-hydroxy-4-cholesten-3-one in peripheral blood
Metabolism Unit, Center for Metabolism and Endocrinology, Department of Medicine and Molecular Nutrition Unit, Center for Nutrition and Toxicology, NOVUM, Karolinska Institute at Huddinge University Hospital, S-141 86 Stockholm, Sweden
1 To whom correspondence should be addressed. e-mail: mats.rudling{at}cnt.ki.se
We describe an accurate method for monitoring the enzymatic activity of hepatic cholesterol 7
-hydroxylase (C7OH; CYP7A1), the rate-limiting and major regulatory enzyme in the synthesis of bile acids. Assay of 7-hydroxy-4-cholesten-3-one (C4), an intermediate in bile acid synthesis, revealed that the level of C4 in peripheral blood serum or plasma showed a strong correlation to the enzymatic activity of hepatic C7OH, both at steady-state conditions (r = 0.929) as well as during the rapid changes that occur during the diurnal phases.
This assay should be of value in clarifying the regulation of bile acid synthesis in vivo in laboratory animals and humans since it allows for the monitoring of hepatic C7OH activity using peripheral blood samples.
Abbreviations: C4, 7-hydroxy-4-cholesten-3-one; C7OH, cholesterol 7-hydroxylase
Supplementary key words CYP7A1 • cholic acid • cholestyramine • high performance liquid chromatography • species
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