Lipid and lipoprotein synthesis in isolated and cultured hepatocytes from lean and obese Zucker rats.
Hepatocytes were isolated by EDTA perfusion of livers from lean (Fa/-) and obese (fa/fa) Zucker rats. Triacylglycerol (TG) and sn-glycerol 3-phosphate were increased in fa/fa hepatocytes, but free fatty acids, cholesterol and phospholipid concentrations were similar in both groups. In spite of an identical fatty acid uptake rate, glycerolipid synthesis was higher in obese compared to lean rat hepatocytes, and this difference remained for at least 2-3 days of culture. Triacylglycerol mass secretion was 2-fold higher in obese than in lean rat hepatocytes. This was confirmed by the higher incorporation of labeled glycerol and oleic acid into the medium TG fraction floating at density 1.006 g/ml. Density gradient ultracentrifugation of [14C]oleate-labeled lipoproteins showed that fa/fa hepatocytes secreted more TG-rich lipoproteins, and that 87% of the label was in the VLDL fraction compared with 67% in the medium of Fa/- hepatocytes. Decreased utilisation of leucine for protein synthesis in obese rat compared to lean rat hepatocytes was associated with enhanced leucine oxidation to CO2. [35S]Methionine incorporation showed an identical cell protein synthesis rate. Autoradiography after PAGE separation of secreted apolipoproteins (apoBh, Bl, apoA-VI, apoE, apoA-I, apoC) showed an identical pattern in both cell types.
Wang
S R
SR
INSERM U.55, Hôpital Saint-Antoine, Paris, France.
Infante
J
J
Catala
D
D
Petit
D
D
Bonnefis
M T
MT
Infante
R
R
D000818:Animals; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003470:Culture Media / Q000032:analysis; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D007930:Leucine / Q000494:pharmacology; D008055:Lipids / Q000096:biosynthesis*; D008074:Lipoproteins / Q000096:biosynthesis*; D008099:Liver / Q000378:metabolism* / Q000648:ultrastructure; D008715:Methionine / Q000494:pharmacology; D019301:Oleic Acid; D009829:Oleic Acids / Q000494:pharmacology; D010168:Palmitates / Q000494:pharmacology; D051381:Rats; D011924:Rats, Zucker
D003470:Culture Media; D008055:Lipids; D008074:Lipoproteins; D009829:Oleic Acids; D010168:Palmitates; D019301:Oleic Acid; D008715:Methionine; D007930:Leucine; D005990:Glycerol