Assay of cholesterol 7α-hydroxylase activity in rat hepatocytes in primary monolayer culture

Princen, H.M.G.; Meijer, Piet; Kwekkeboom, Jaap; Kempen, H.J.M.

doi:10.1016/0003-2697(88)90137-6

Cited by 26 publications

(36 citation statements)

References 31 publications

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“…Cholesterol 7IX-hydroxylase activity in rat hepatocytes in primary monolayer culture was measured by the method of Princen et al 10 ) The cells (5 x 10 6 ) seeded on culture dishes (100 x lOmm, FALCON) were incubated for 48 h with 10 ml of medium as described above. The reaction was stopped by the addition of a chloroform-methanol mixture (2: 1, vJv) containing butylated hydroxytoluene.…”

Section: Methodsmentioning

confidence: 99%

Propionate Enhances Synthesis and Secretion of Bile Acids in Primary Cultured Rat HepatocytesviaSuccinyl CoA

Imaizumi

Hirata

Yasni

et al. 1992

Bioscience, Biotechnology, and Biochemistry

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Section: Methodsmentioning

confidence: 99%

Propionate Enhances Synthesis and Secretion of Bile Acids in Primary Cultured Rat HepatocytesviaSuccinyl CoA

Imaizumi

Hirata

Yasni

et al. 1992

Bioscience, Biotechnology, and Biochemistry

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“…Rat hepatocyte preparation and culture Rat liver cells were isolated by perfusion with 0.05 % collagenase and 0.0050% trypsin inhibitor as described previously [19][20][21]. Viability, as determined by Trypan Blue exclusion, was higher than 90 %.…”

Section: Materials and Animalsmentioning

confidence: 99%

“…Materials used for the isolation and culture of rat and human hepatocytes, determination of bile acid synthesis from radiolabelled cholesterol, measurement of mass production of bile acids, the cholesterol 7a-hydroxylase activity and other enzyme assays were obtained from sources described previously [19][20][21].…”

Section: Materials and Animalsmentioning

confidence: 99%

“…Cholesterol 70c-hydroxylase and mitochondrial 26-hydroxylase assays Cholesterol 7a-hydroxylase activity was measured as reported in microsomes [19] or in homogenates of freshly isolated hepatocytes [20]. 26 [27] and Gamble et al [28] respectively.…”

Section: Materials and Animalsmentioning

confidence: 99%

“…The cells were seeded on 6-well cluster plates or on 60 mm-diam. plastic tissue culture dishes (Costar, Cambridge, MA, U.S.A.) at a density of 1 x 105 cells/cm2 and were maintained in Williams E medium supplemented with 10 % (v/v) heat-inactivated foetal bovine serum, 2 mM-L-glutamine, 20 munits of insulin/ml, 50 nM-dexamethasone, 100 units of penicillin/ml and 100 ug of streptomycin/ml at 37°C in an atmosphere of air/CO2 (19:1) [19,20]. After a 4 h attachment period, and every 24 h thereafter, medium was replaced with I ml (wells) or 2.5 ml (dishes) of culture medium.…”

Section: Materials and Animalsmentioning

confidence: 99%

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Cyclosporin A blocks bile acid synthesis in cultured hepatocytes by specific inhibition of chenodeoxycholic acid synthesis

Princen¹,

Meijer

Wolthers

et al. 1991

Biochemical Journal

102

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Bile acid synthesis, determined by conversion of [4-14C]cholesterol into bile acids in rat and human hepatocytes and by measurement of mass production of bile acids in rat hepatocytes, was dose-dependently decreased by cyclosporin A, with 52 % (rat) and 45 % (human) inhibition at 10/,M. The decreased bile acid production in rat hepatocytes was due only to a fall in the synthesis of,-muricholic and chenodeoxycholic acids (-64 % at 10 4M-cyclosporin A), with no change in the formation of cholic acid. In isolated rat liver mitochondria, 26-hydroxylation of cholesterol was potently inhibited by the drug (concn. giving half-maximal inhibition = 4/tM). These results suggest that cyclosporin A blocks the alternative pathway in bile acid synthesis, which leads preferentially to the formation of chenodeoxycholic acid.

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Lifibrol increases hepatic cholesterol 7α‐hydroxylase activity in sprague‐dawley rats

Dinh¹,

Funk

Vidmar³

et al. 1994

Drug Development Research

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The hypocholesterolemic drug lifibrol was administered orally to Sprague-Dawley rats to determine its effects on lipoprotein cholesterol distribution and hepatic HMG-CoA reductase and cholesterol 7a-hydroxylase activities. The effects of lifibrol on those endpoints were compared with the effects of gemfibrozil and lovaytatin. When administered to either chow-fed or cholesterol-fed rats, lifibrol (25 or 50 mglkgiday) caused a redistribution of lipoprotein cholesterol such that HDL increased and VLDL+ LDL decreased significantly Gemfibroril (30 or 50 mgikglday) caused similar changes in lipoprotein cholesterol distribution. In contrast, lovastatin (10 mgikglday) decreased both HDL and VLDL+LDL in chow-fed animals, but had no effect in cholesterol-fed animals. Hepatic HMG-CoA reductase activity was increased in rats treated with lifibrol (50 rnglkgl day), gemfibrozil (50 mglkgiday), and lovastatin (10 mgikgiday). 1 he most significant finding is that lifibrol was the only drug that increased hepatic cholesterol 7a-hydroxylase activity. This observation in rats separates the mechanism of action of lifibrol from those of the HMG-CoA reductase inhibitors and the fibrates, and suggests that one mechanism ot action of litibrol is to enhance cholesterol elimination through conversion to bile acids.1994 b\'iley ~i s s Inc

show abstract

Assay of cholesterol 7α-hydroxylase activity in rat hepatocytes in primary monolayer culture

Cited by 26 publications

References 31 publications

Propionate Enhances Synthesis and Secretion of Bile Acids in Primary Cultured Rat HepatocytesviaSuccinyl CoA

Propionate Enhances Synthesis and Secretion of Bile Acids in Primary Cultured Rat HepatocytesviaSuccinyl CoA

Cyclosporin A blocks bile acid synthesis in cultured hepatocytes by specific inhibition of chenodeoxycholic acid synthesis

Lifibrol increases hepatic cholesterol 7α‐hydroxylase activity in sprague‐dawley rats

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