Origin of hepatic very-low-density lipoprotein triacylglycerol: The contribution of cellular phospholipid

When rat hepatocytes were cultured for 24 h in the absence of exogenous fatty acid, the amount of very-low-density lipoprotein (VLDL) triacylglycerol (TAG) secreted (114 +/- 14 mu g/mg of cell protein) could not be accounted for by the mass of TAG lost from the cells (29 +/- 6.1 mu g/mg of cell protein) during this period (n = 12). Of the balance (85 +/- 14 mu g/mg; 94 +/- 15 nmol/mg), a maximum of only 37 nmol/mg of cell protein of TAG could be accounted for by fatty acids synthesized de novo. When labelled exogenous oleate (initial concentration, 0.75 mM) was present in the culture medium, the net gain in cellular plus VLDL TAG (253 +/- 38 mu g/mg of cell protein per 24 h) was greater than that contributed by the exogenous fatty acid (155 +/- 18.2 mu g/mg of cell protein, n = 5). Again, the balance (98.8 +/- 18.2 mu g/mg of cell protein per 24 h) was too great to be accounted for by fatty acid synthesis de nova. In experiments in which cellular glycerolipids were prelabelled with [9,10(n)-H-3]oleic acid, following removal of the labelled fatty acid, there was a net increase in labelled cellular plus VLDL TAG over the next 24h. That cellular phospholipids are the source of a substantial part of the excess TAG synthesized is supported by the following evidence. (1) The loss of prelabelled cellular phospholipid during culture was greater than could be accounted for by secretion into the medium. (2) During culture of cells prelabelled with 1,2-di-[C-114]palmitoyl phosphatidylcholine, a substantial amount of label was secreted as VLDL TAG. (3) In pulse-chase experiments, the kinetics of labelled phospholipid turnover were consistent with conversion into a non-phospholipid pool. The enzymology involved in the transfer of phospholipid fatty acids into TAG is probably complex, but the present results suggest that this pathway may represent an important route by which extracellular fatty acids are channelled into VLDL TAG.