Dissimilar effects of Brefeldin A on cholesteryl ester and triacylglycerol metabolism in CaCo2 and HepG2 cells as compared to peritoneal macrophages.
The effect of Brefeldin A (BFA) on lipid metabolism was studied in two cell lines and in primary cultures of peritoneal macrophages. In both CaCo2 and HepG2 cells, which are models for human liver and intestine, addition of BFA resulted in a 2-10-fold increase in recovery of labeled cholesteryl ester when the cells had been prelabeled with free cholesterol or with [3H]oleic acid. This effect was linear for up to 6 h and could be elicited with doses of BFA as low as 0.03 micrograms/ml. The increase in cholesteryl ester induced by BFA was completely abolished by the ACAT inhibitor (Sandoz 58-035) and partly by forskolin. Intracellular hydrolysis of labeled cholesteryl ester was studied in the presence of the ACAT inhibitor and while in the controls 30-40% was hydrolyzed in 6 h, the values were 7-16% in the BFA treated cells. The slower rate of hydrolysis in the BFA treated cells could not account for the entire increase of cholesteryl ester and there was also no decrease in cholesteryl ester secretion. Even though activation of acyl-CoA:cholesterol acyltransferase by BFA was not demonstrated in cell homogenates, we hypothesize that in the intact cell the BFA induced increase in cholesteryl ester might have been related to the pronounced increase in modified endoplasmic reticulum which results from the dispersion of the Golgi apparatus. In the macrophages, BFA at doses of 0.25-1 micrograms/ml resulted in a 90% reduction in the incorporation of [3H]oleic acid into triacyglycerol. Incorporation of [3H]oleic acid into triacyglycerol in CaCo2 cells was not affected by BFA. In view of the ever-increasing use of BFA in cell biology, it seems important to emphasize that BFA may affect different pathways of lipid metabolism in various cells.
Stein
O
O
Department of Experimental Medicine and Cancer Research, Hebrew University-Hadassah Medical School Jerusalem, Israel.
Dabach
Y
Y
Hollander
G
G
Ben-Naim
M
M
Stein
Y
Y
D000107:Acetylation; D020126:Brefeldin A; D002460:Cell Line; D002788:Cholesterol Esters / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology*; D004951:Esterification / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D009829:Oleic Acids / Q000378:metabolism; D010537:Peritoneum; D014280:Triglycerides / Q000378:metabolism*
D002788:Cholesterol Esters; D003517:Cyclopentanes; D009829:Oleic Acids; D014280:Triglycerides; D020126:Brefeldin A