The objective of these experiments was to investigate the source of substrates used for lipid synthesis and the pathways of substrate incorporation into lipids by epithelial cells of the colon. Within replicates, cells were exposed to all treatments evaluated in that experiment. By comparing the relative incorporation rates of several 14C-labeled substrates into lipids, acetate made a significantly larger carbon contribution to lipids than propionate, butyrate, glucose or glutamine under the in vitro conditions utilized in this study. Other major carbon contributors were butyrate and 3-hydroxybutyrate. Glucose, glutamine and propionate made only minor contributions. (-)-Hydroxycitrate did not affect the incorporation of acetate or butyrate carbon into lipids, even though it inhibited colonic ATP-citrate lyase. These data suggest that SCFA carbon used in the synthesis of lipids by colonocytes is not likely transported to the cytosol as citrate. Competition experiments suggest that ketone bodies and butyrate contribute to a single precursor pool for lipogenesis. Ketone bodies did not significantly suppress acetate incorporation into lipid, however. Incorporation of 3H2O and 14C-acetate was significantly greater into phospholipids than into free fatty acids and triacylglycerides, suggesting that the major role of lipogenesis is for membrane synthesis. In conclusion, colonocytes appear to synthesize lipids using a pathway distinct from the liver by incorporating mainly SCFA and ketone bodies into lipids, and by using citrate to a limited extent, if at all, to transport acetyl units from the mitochondria to the cytosol.
