Home LiteratureArticle Details
PMID: 8687397 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of the inhibitory effects of bile acids on very-low-density lipoprotein secretion by rat hepatocytes in primary culture.

The Biochemical journal ·Vol. 316 ( Pt 2) ·1996-06-01 ·Pages 531-8

Lin Y, Havinga R, Schippers IJ, Verkade HJ, Vonk RJ, Kuipers F

Abstract

In this study the effects of bile acids and other organic anions on the secretion of very-low-density lipoproteins (VLDLs) were evaluated in rat hepatocytes in primary culture. Incubation of cells with portal blood concentrations (10-200 microM) of bile acids resulted in dose-dependent suppression of secretion of VLDL-associated [3H]triglyceride (TG) formed from [3H]glycerol, and also of TG mass. The degree of the inhibition was highly correlated with intracellular bile acid concentration. Prolonged incubation with 100 microM extracellular taurocholic acid (TC) decreased the secretion of [3H]TG and TG mass to 35% and 50% of the controls respectively. Cellular content of mass and of [3H]TG during prolonged incubation with TC were about 20% and 60% higher than the controls respectively. The inhibitory effect remained for at least 24 h in the presence of TC without altering VLDL-lipid and VLDL-apolipoprotein compositions or the size distribution of the particles. Secretion of apoB-100 and of apoB-48 was inhibited to a similar extent. Cells largely lost their capacity to accumulate bile acids intracellularly after 48 h in culture. In these cells TC was unable to exert its suppressive effects. Taurine and glycine conjugates of all common bile acids were capable of suppressing [3H]TG secretion. Trihydroxylated (cholic acid) and various dihydroxylated (deoxycholic, chenodeoxycholic and ursodeoxycholic acids) bile acids had similar capacities in this respect, suggesting that their common sterol-3 alpha-OH structure is required for the suppressive effect. Neither non-bile acid organic anions, e.g. bilirubin ditaurate and dibromosulphthalein, nor dianionic bile acid metabolites, e.g. sulphated taurolithocholic acid and lithocholate-3-O-glucuronide, showed any effect on [3H]TG secretion. These results indicate that bile acids might play a physiological role in regulating VLDL production by the liver, especially in the postprandial state when their enterohepatic circulation is stimulated.

MeSH Terms
Animals Apolipoproteins B/metabolism Bile Acids and Salts/pharmacology Cells, Cultured Cholic Acids/pharmacology Lipoproteins, VLDL/metabolism Liver/cytology,metabolism Male Particle Size Proteins/metabolism Rats Rats, Wistar Serum Albumin/metabolism Taurocholic Acid/metabolism,pharmacology Triglycerides/metabolism
Chemicals
Apolipoproteins B Bile Acids and Salts Cholic Acids Lipoproteins, VLDL Proteins Serum Albumin Triglycerides Taurocholic Acid
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lin Y
Groningen Institute for Drug Studies, Department of Paediatrics, Academic Hospital Groningen, The Netherlands.
Havinga R
Schippers I J
Verkade H J
Vonk R J
Kuipers F
References (46)
46 references, click to expand
  1. Bile acids in portal blood of fats fed different diets and cholestyramine. Bile acids and steroids 189.
    Eur J Biochem. 1967 Nov;2(4):375-83 PMID: 6080771
  2. Secretion of biliary lipids from the hepatocyte.
    Hepatology. 1990 Sep;12(3 Pt 2):134S-141S; discussion 141S-142S PMID: 2210641
  3. The binding of detergents to lipophilic and hydrophilic proteins.
    J Biol Chem. 1972 Jun 10;247(11):3656-61 PMID: 4113126
  4. Dynamics of the enterohepatic circulation of bile acids. Postprandial serum concentrations of conjugates of cholic acid in health, cholecystectomized patients, and patients with bile acid malabsorption.
    N Engl J Med. 1974 Oct 3;291(14):689-92 PMID: 4851463
  5. Synthesis and secretion of rat albumin in vivo, in perfused liver, and in isolated hepatocytes. Effects of hypophysectomy and growth hormone treatment.
    J Biol Chem. 1977 Jun 10;252(11):3611-6 PMID: 863898
  6. Postprandial serum bile acids in healthy man. Evidence for differences in absorptive pattern between individual bile acids.
    Gut. 1977 Aug;18(8):606-9 PMID: 892604
  7. Conjugates of ursodeoxycholate protect against cholestasis and hepatocellular necrosis caused by more hydrophobic bile salts. In vivo studies in the rat.
    Gastroenterology. 1991 Jan;100(1):203-11 PMID: 1983822
  8. Regulation of apoB secretion from HepG2 cells: evidence for a critical role for cholesteryl ester synthesis in the response to a fatty acid challenge.
    J Lipid Res. 1990 Nov;31(11):2045-55 PMID: 1964953
  9. Prostaglandins suppress VLDL secretion in primary rat hepatocyte cultures: relationships to hepatic calcium metabolism.
    J Lipid Res. 1992 Jul;33(7):1017-27 PMID: 1331281
  10. Effects of tauroursodeoxycholic acid on cytosolic Ca2+ signals in isolated rat hepatocytes.
    Gastroenterology. 1993 Feb;104(2):604-12 PMID: 8425704
  11. Substrate delivery as a determinant of hepatic apoB secretion.
    Arterioscler Thromb. 1993 May;13(5):629-36 PMID: 8485114
  12. Comparative binding of bile acids to serum lipoproteins and albumin.
    J Lipid Res. 1993 Oct;34(10):1661-74 PMID: 8245717
  13. Deficiency of bile acid transport and synthesis in oval cells from carcinogen-fed rats.
    Hepatology. 1994 Mar;19(3):722-7 PMID: 8119699
  14. Apparent selective bile acid malabsorption as a consequence of ileal exclusion: effects on bile acid, cholesterol, and lipoprotein metabolism.
    Gut. 1994 Aug;35(8):1116-20 PMID: 7926917
  15. Differential effects of eicosapentaenoic acid on glycerolipid and apolipoprotein B metabolism in primary human hepatocytes compared to HepG2 cells and primary rat hepatocytes.
    Biochim Biophys Acta. 1995 Apr 28;1256(1):88-96 PMID: 7742360
  16. Effects of ursodeoxycholic acid and chenodeoxycholic acid on human hepatocytes in primary culture.
    Hepatology. 1995 Jul;22(1):82-7 PMID: 7601437
  17. Bile acids suppress the secretion of very-low-density lipoprotein by human hepatocytes in primary culture.
    Hepatology. 1996 Feb;23(2):218-28 PMID: 8591844
  18. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  19. Phosphorus assay in column chromatography.
    J Biol Chem. 1959 Mar;234(3):466-8 PMID: 13641241
  20. A rapid method of total lipid extraction and purification.
    Can J Biochem Physiol. 1959 Aug;37(8):911-7 PMID: 13671378
  21. Individual bile acids in portal venous and systemic blood serum of fasting man.
    Gastroenterology. 1977 Dec;73(6):1377-82 PMID: 913977
  22. Physiological factors influencing serum bile acid levels.
    Gut. 1978 Jan;19(1):32-9 PMID: 624503
  23. Increased production of triglyceride-rich lipoproteins after partial biliary diversion in the rhesus monkey.
    Metabolism. 1978 May;27(5):607-13 PMID: 417230
  24. Effects of cholestyramine and chenodeoxycholic acid on the metabolism of endogenous triglyceride in hyperlipoproteinemia.
    J Lipid Res. 1978 Nov;19(8):1017-24 PMID: 731123
  25. Procedure for determination of free and total cholesterol in micro- or nanogram amounts suitable for studies with cultured cells.
    J Lipid Res. 1978 Nov;19(8):1068-70 PMID: 731127
  26. Very low density lipoprotein synthesis and secretion by cultured rat hepatocytes.
    J Biol Chem. 1979 Mar 25;254(6):2010-6 PMID: 217875
  27. Improved method for accurate quantitation of total and conjugated bilirubin in serum.
    Clin Chem. 1979 Nov;25(11):1891-9 PMID: 40710
  28. Effects of cholic acid on the metabolism of endogenous plasma triglyceride and on biliary lipid composition in hyperlipoproteinemia.
    J Lipid Res. 1980 Jan;21(1):1-9 PMID: 7354246
  29. Isolation and analysis of lipoproteins secreted by rat liver hepatocytes.
    Biochim Biophys Acta. 1984 Jan 17;792(1):39-47 PMID: 6419782
  30. Influence of hydroxylation and conjugation of bile salts on their membrane-damaging properties--studies on isolated hepatocytes and lipid membrane vesicles.
    Hepatology. 1984 Jul-Aug;4(4):661-6 PMID: 6745854
  31. Suitability of primary monolayer cultures of adult rat hepatocytes for studies of cholesterol and bile acid metabolism.
    J Biol Chem. 1985 Jan 25;260(2):1015-9 PMID: 3968056
  32. Electron microscopy of negatively stained lipoproteins.
    Methods Enzymol. 1986;128:442-57 PMID: 2425222
  33. Ursodeoxycholic acid treatment in humans: effects on plasma and biliary lipid metabolism with special reference to very low density lipoprotein triglyceride and bile acid kinetics.
    Eur J Clin Invest. 1986 Apr;16(2):169-77 PMID: 3089812
  34. Ketoconazole blocks bile acid synthesis in hepatocyte monolayer cultures and in vivo in rat by inhibiting cholesterol 7 alpha-hydroxylase.
    J Clin Invest. 1986 Oct;78(4):1064-71 PMID: 3760182
  35. Eicosapentaenoic acid inhibits synthesis and secretion of triacylglycerols by cultured rat hepatocytes.
    Biochim Biophys Acta. 1986 Oct 24;879(1):56-65 PMID: 3021226
  36. The active synthesis of phosphatidylcholine is required for very low density lipoprotein secretion from rat hepatocytes.
    J Biol Chem. 1988 Feb 25;263(6):2998-3004 PMID: 3343237
  37. Bile protein secretion in the rat stimulated by taurocholate: effect of chloroquine.
    Can J Physiol Pharmacol. 1988 Jun;66(6):749-53 PMID: 3167691
  38. Immunoperoxidase localization of bile salts in rat liver cells. Evidence for a role of the Golgi apparatus in bile salt transport.
    J Clin Invest. 1988 Oct;82(4):1173-82 PMID: 2459158
  39. Regulation of biliary cholesterol secretion. Functional relationship between the canalicular and sinusoidal cholesterol secretory pathways in the rat.
    J Clin Invest. 1988 Dec;82(6):1818-25 PMID: 3198756
  40. Comparison of taurocholate accumulation in cultured hepatocytes of pig, rat and man.
    Biochem Biophys Res Commun. 1989 Jul 31;162(2):619-25 PMID: 2757635
  41. Chenodeoxycholic acid normalizes elevated lipoprotein secretion and catabolism in cerebrotendinous xanthomatosis.
    J Lipid Res. 1989 May;30(5):633-40 PMID: 2760539
  42. Lipid and carbohydrate metabolism in cholestyramine-fed rats.
    Artery. 1989;16(5):286-92 PMID: 2764732
  43. Regulation of bile acid synthesis. III. Correlation between biliary bile salt hydrophobicity index and the activities of enzymes regulating cholesterol and bile acid synthesis in the rat.
    J Lipid Res. 1989 Aug;30(8):1161-71 PMID: 2769071
  44. Insulin modulation of hepatic synthesis and secretion of apolipoprotein B by rat hepatocytes.
    J Biol Chem. 1990 May 25;265(15):8854-62 PMID: 2187873
  45. Ursodeoxycholate reduces hepatotoxicity of bile salts in primary human hepatocytes.
    Hepatology. 1990 Sep;12(3 Pt 1):486-91 PMID: 2401454
  46. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study.
    J Cell Biol. 1969 Dec;43(3):506-20 PMID: 4900611
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
Published
1996-06-01
Pages
531-8
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1217381
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]