Monitoring hepatic cholesterol 7alpha-hydroxylase activity by assay of the stable bile acid intermediate 7alpha-hydroxy-4-cholesten-3-one in peripheral blood.
We describe an accurate method for monitoring the enzymatic activity of hepatic cholesterol 7alpha-hydroxylase (C7alphaOH; CYP7A1), the rate-limiting and major regulatory enzyme in the synthesis of bile acids. Assay of 7alpha-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 C7alphaOH, 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 C7alphaOH activity using peripheral blood samples.
Gälman
Cecilia
C
Metabolism Unit, Center for Metabolism and Endocrinology, S-141 86 Stockholm, Sweden.
Arvidsson
Ingela
I
Angelin
Bo
B
Rudling
Mats
M
D000818:Animals; D001647:Bile Acids and Salts / Q000096:biosynthesis*; D002783:Cholestenones / Q000097:blood*; D002790:Cholesterol 7-alpha-Hydroxylase / Q000032:analysis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002940:Circadian Rhythm; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008297:Male; D008722:Methods; D051381:Rats; D017208:Rats, Wistar
D001647:Bile Acids and Salts; D002783:Cholestenones; C002656:7 alpha-hydroxy-4-cholesten-3-one; D002790:Cholesterol 7-alpha-Hydroxylase