title: Evaluation of cultured hamster hepatocytes as an experimental model for the study of very low density lipoprotein secretion.
abstract: The secretion of triacylglycerol, cholesterol and cholesteryl ester in very low density lipoprotein (VLDL) by cultured hamster hepatocytes was studied, and the results compared with those obtained previously using cultured rat hepatocytes and the human hepatoma cell line HepG2. The hamster cells secreted apolipoprotein B and VLDL triacylglycerol, cholesterol and cholesteryl ester linearly during 24 h in culture, and this time period was used in all experiments. Addition of oleate (1 mM) to the culture medium resulted in increased secretion of triacylglycerol, but cholesterol ester output were unchanged. Triacylglycerol secretion was also increased in the presence of lipogenic substrates (10 mM lactate + 1 mM pyruvate) plus dexamethasone (1 microM), but not with either of these agents alone. Inhibition of cholesterol synthesis in the hamster cells by incubation with mevinolin (2 micrograms/ml) did not change VLDL lipid secretion, but stimulation using mevalonate lactone resulted in decreased triacylglycerol output. Manipulation of the rate of cholesterol esterification in the hepatocytes by inhibiting or stimulating the activity of acyl coenzyme A cholesterol:acyl transferase using the inhibitor Dup128 (25 microM) and 25-hydroxycholesterol (50 microM), respectively, had no effect on the secretion of VLDL lipid. In the presence of 1 mM oleate plus 25-hydroxycholesterol, however, a rise in the output of triacylglycerol and cholesteryl ester was observed. Hepatocytes prepared from hamsters fed 2% cholestyramine secreted significantly less triacylglycerol than those from animals given the control diet, but cholesterol and cholesteryl ester output were unchanged, despite a decrease of about 40% in the total cholesterol content of the cells. These results show that the secretion of lipid in VLDL in hamster hepatocytes differs from that in rat and human liver in its response to dietary cholestyramine, and from rat hepatocytes and HepG2 cells in its response to changes in the rate of lipogenesis and cholesterol synthesis and esterification. Overall, hamster hepatocytes appear to be less susceptible to modification the rate of hepatic VLDL secretion, and should provide a useful additional tool for the investigation of this process.
authors: Department of Veterinary Basic Sciences, Royal Veterinary College, London, UK.
authors: V Q
authors: VQ
authors: Hoang
authors: N J
authors: NJ
authors: Pearce
authors: K E
authors: KE
authors: Suckling
authors: K M
authors: KM
authors: Botham
mesh_terms: D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology; D002478:Cells, Cultured; D002784:Cholesterol / Q000032:analysis; D002788:Cholesterol Esters / Q000032:analysis; D002791:Cholesterol, Dietary / Q000494:pharmacology; D006224:Cricetinae; D003907:Dexamethasone / Q000494:pharmacology; D007773:Lactates / Q000494:pharmacology; D019344:Lactic Acid; D008079:Lipoproteins, VLDL / Q000096:biosynthesis*; D008099:Liver / Q000378:metabolism*; D008148:Lovastatin / Q000494:pharmacology; D008297:Male; D008647:Mesocricetus; D008954:Models, Biological; D019301:Oleic Acid; D009829:Oleic Acids / Q000494:pharmacology; D011773:Pyruvates / Q000494:pharmacology; D019289:Pyruvic Acid; D014280:Triglycerides / Q000032:analysis
chemical_list: D000924:Anticholesteremic Agents; D002788:Cholesterol Esters; D002791:Cholesterol, Dietary; D007773:Lactates; D008079:Lipoproteins, VLDL; D009829:Oleic Acids; D011773:Pyruvates; D014280:Triglycerides; D019301:Oleic Acid; D019344:Lactic Acid; D003907:Dexamethasone; D019289:Pyruvic Acid; D002784:Cholesterol; D008148:Lovastatin