orlistat and cholesteryl-oleate

Though it is now clear that chylomicron remnants are pro-atherogenic lipoproteins, events leading to their incorporation by macrophages are poorly understood.. This study investigates, in human macrophages, the fate of either [(3)H]cholesteryl oleate or [(3)H]triacylglycerol carried by human apolipoprotein-E-containing chylomicron remnant-like particles (CRLP) and the influence of CRLP containing trilinolein, (18:2)CRLP, or triolein, (18:1)CRLP, on lipid accumulation, newly synthesized cholesteryl ester (CE) and triacylglycerol (TG).. Labelled fatty acids from TG were markedly incorporated into TG and phospholipid and, to a lesser extent, into free fatty acids and were scarcely recovered in cholesteryl esters. [(3)H]CE from CRLP accumulated in cells in a dose-dependent manner with a significant difference between concentrations of 10 and 40 microg cholesterol/ml with (18:2)CRLP. In the same concentration range, TG synthesis was enhanced by about 46 and 30% by (18:2)CRLP and (18:1)CRLP cholesterol, respectively, whereas the esterification of cholesterol, evaluated by [(3)H]oleate incorporation, was decreased by about 30% with both types of CRLP. Endocytosis inhibition did not prevent cell cholesterol and TG accumulation, whereas lipoprotein lipase inhibition reduced the TG content.. The results are consistent with the hypotheses that in macrophages dietary remnants may support TG and CE internalization via different mechanisms. Extracellular lipolysis seems particularly important for internalization of dietary fatty acids, whereas the entrance of CE seems attributable to a concomitant selective CE uptake mediated by scavenger receptor class B type I, since the scavenger receptor class B type I antibody induces significant inhibition (38%) of [(3)H]CE transported by CRLP, but does not affect internalization of [(3)H]TG carried by the same particles.

Topics: Antibodies, Monoclonal; Apolipoproteins E; Biological Transport; Cells, Cultured; Cholesterol Esters; Chylomicron Remnants; Chylomicrons; Enzyme Inhibitors; Fatty Acids, Nonesterified; Humans; Lactones; Lipoprotein Lipase; Macrophages; Orlistat; Particle Size; Phospholipids; Scavenger Receptors, Class B; Triglycerides; Tritium

Tetrahydrolipstatin (THL) derived by hydrogenation from lipstatin, a lipase inhibitor produced by Streptomyces toxytricini, has been shown to inhibit in vitro the activity of all three lipases secreted to the gastro-intestinal tract; gastric lipase, pancreatic lipase and carboxylester lipase (cholesterol ester hydrolase). The effects of THL on intestinal absorption of fat (transport to the thoracic duct chyle) has now been investigated after intraduodenal infusion in a rat model. Absorption of label from oleic acid when administered with monoolein in micellar bile salt solution was not affected by THL in concentrations up to 10(-4) M calculated on the volume of the aqueous phase. Absorption of free cholesterol in micellar bile salt solution of the lipolytic products of triolein; oleic acid and monoolein, is not significantly affected at a concentration of THL of 10(-4) M. Absorption of cholesterol from cholesteryl oleate under the same conditions is almost completely inhibited. The results indicate that absorption of free cholesterol is not dependent on the activity of pancreatic cholesterol ester hydrolase. The absorption of emulsified triolein was not significantly affected by 10(-5) M THL but decreased to around 30% of the controls by a concentration 10-times higher. There was no significant decrease of cholesterol absorption when administered in emulsified triolein while absorption of cholesteryl oleate was reduced at both concentrations of THL and almost completely at 10(-4) M. Radioactivity from [2-14C]THL when administered emulsified in triolein was recovered in urine, bile and thoracic duct lymph to 10-14, 8-13 and 1-3%, respectively, largely independent on dose administered. Label from [1"-14C] THL was recovered in the same amounts in lymph but much less in bile and urine indicating that the amino acid moiety has been split off early in the absorption process.

Topics: Animals; Cholesterol; Cholesterol Esters; Dietary Fats; Emulsions; Intestinal Absorption; Lactones; Lipase; Male; Micelles; Oleic Acid; Oleic Acids; Orlistat; Rats; Rats, Inbred Strains