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40
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Text
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Pages
16866-16871
Issue
24
Volume
267
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Title
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Hormonal-regulation Of Cholesterol-7-alpha-hydroxylase Messenger-rna Levels And Transcriptional Activity In Primary Rat Hepatocyte Cultures
Publisher
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Journal of Biological Chemistry
Date
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1992
1992-08
Subject
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bile-acid synthesis; Biochemistry & Molecular Biology; chick-embryo hepatocytes; cholesterol 7-alpha-hydroxylase activity; circadian-rhythm; cyclic-amp; enzyme; gene; liver; malic; metabolism; monolayer-cultures
Creator
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Hylemon P B; Gurley E C; Stravitz R T; Litz J S; Pandak W M; Chiang J Y L; Vlahcevic Z R
Description
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In primary cultures of adult rat hepatocytes the level of cholesterol 7-alpha-hydroxylase steady-state mRNA markedly decreased by 72 h. However, the addition of L-thyroxine (T4) and dexamethasone synergistically returned cholesterol 7-alpha-hydroxylase steady-state mRNA levels near to that of cholestyramine-fed animals. The maximal responses to T4 and dexamethasone in serum-free medium were at 1.0 and 0.1-mu-M, respectively. The addition of T4 in combination with dexamethasone resulted in an 11-fold increase in transcriptional activity of the cholesterol 7-alpha-hydroxylase gene as compared to no addition controls. The specific activities of cholesterol 7-alpha-hydroxylase in microsomes prepared from cultures treated with dexamethasone and T4 were 1.56 +/- 1.17 nmol/h/mg protein which is similar to that of intact liver (1.70 +/- 0.062 nmol/h/mg protein), but lower than cholestyramine-fed animals. Cholesterol 7-alpha-hydroxylase activity was not detectable (<0.020 nmol/h/mg protein) at 72 h in cultures without the addition of both dexamethasone and T4. In the presence of optimal concentrations of dexamethasone and T4, glucagon (0.2-mu-M), or dibutyryl cAMP (50-mu-M) decreased (90%) cholesterol 7-alpha-hydroxylase mRNA within 6 h. Transcriptional activity decreased (62%) in 6 h following the addition of glucagon (0.2-mu-M) to the culture medium. The results reported in this paper suggest an important role for multiple hormones in the regulation of cholesterol 7-alpha-hydroxylase in the liver.
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n/a
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Journal Article or Conference Abstract Publication
1992
bile-acid synthesis
Biochemistry & Molecular Biology
Chiang J Y L
chick-embryo hepatocytes
cholesterol 7-alpha-hydroxylase activity
circadian-rhythm
cyclic-amp
enzyme
gene
Gurley E C
Hylemon P B
Journal Article or Conference Abstract Publication
Journal of Biological Chemistry
Litz J S
Liver
malic
Metabolism
monolayer-cultures
Pandak W M
Stravitz R T
Vlahcevic Z R