Effects of albumin on lipid synthesis, apo B-100 secretion, and LDL catabolism in HepG2 cells.
This study examines the effects of extracellular albumin on hepatic apo B-100 metabolism. To do so, a transformed human liver cell line, HepG2, was used as a hepatocyte model and the concentration of albumin in the medium was varied between 0 and 5 g%. Apo B-100 and apo A1 concentrations in the medium were determined by specific enzyme-linked immunoassay (ELISA) and intracellular synthesis of cholesterol ester and triglyceride were determined by addition of appropriate radiolabels to the medium. The data demonstrate that the reduction of extracellular albumin concentration resulted in increased apo B-100 concentration in the medium. Apo A1 secretion, however, was unaffected. While the differences in apo B-100 concentration in the medium were statistically significant (33% +/- 7%, P < 0.0025, 0 g% albumin compared to 5 g% albumin in the medium), the absolute magnitude of the effect under these conditions was relatively modest. Nevertheless, the changes were consistent and evident over incubation periods as long as 8 days. Of interest, although triglyceride synthesis was unaffected, cholesterol ester synthesis changed such that as albumin concentration decreased, synthesis of cholesterol ester increased paralleling the changes in apo B-100 (170% +/- 9%, P < 0.005). These findings were extended by studying interventions which altered cholesterol ester synthesis. Addition of the compound 58-035 (5 micrograms/ml, a specific inhibitor of acylcholesterol acyltransferase activity) resulted in substantial inhibition of cholesterol ester synthesis (39% to 66%, P < 0.025 and P < 0.005, respectively) and apo B-100 concentrations in the medium which decreased by 20% to 28%, P < 0.025. Triglyceride synthesis, in contrast, increased significantly by 32% P < 0.025. Therefore, addition of 58-035 confirmed the previous findings of a parallel relation between cholesterol ester synthesis and apo B-100 concentration in the medium. Nonetheless, albumin still had an additional inhibitory effect on cholesterol ester and apo B-100 secretion. Of interest, when chylomicron remnants (25 micrograms/ml cholesterol), which cause apo B-100 secretion to increase by more than threefold, were added to the medium, albumin now had a more pronounced absolute effect on apo B-100 secretion with a 48% inhibition observed as albumin was increased from 0 to 5 g% in the medium (P < 0.0125). The effect of extracellular albumin on the low density lipoprotein (LDL) pathway was also examined. No differences in non-specific cell association component were detected.(ABSTRACT TRUNCATED AT 400 WORDS)
Cianflone
K
K
McGill Unit for the Prevention of Cardiovascular Disease, Royal Victoria Hospital, Montreal, Canada.
Vu
H
H
Zhang
Z
Z
Sniderman
A D
AD
D000577:Amides / Q000494:pharmacology; D016632:Apolipoprotein A-I / Q000378:metabolism; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000378:metabolism*; D002461:Cell Line, Transformed; D002784:Cholesterol / Q000096:biosynthesis; D002788:Cholesterol Esters / Q000096:biosynthesis; D002914:Chylomicrons / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008055:Lipids / Q000096:biosynthesis*; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D017646:Organosilicon Compounds; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D002785:Sterol O-Acyltransferase / Q000037:antagonists & inhibitors; D014280:Triglycerides / Q000096:biosynthesis
D000577:Amides; D016632:Apolipoprotein A-I; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D002788:Cholesterol Esters; D002914:Chylomicrons; D008055:Lipids; D008077:Lipoproteins, LDL; D017646:Organosilicon Compounds; D014280:Triglycerides; D012710:Serum Albumin, Bovine; C040256:SAN 58035; D002784:Cholesterol; D002785:Sterol O-Acyltransferase