We explored potential mechanisms of non-low-density lipoprotein (LDL) receptor-mediated uptake of triglyceride-rich particles (TGRP) in the presence of apolipoprotein E (apo E). Human fibroblasts were incubated with model intermediate-density lipoprotein- (IDL-) sized TGRP (10-1000 microg of neutral lipid/mL) containing apo E. The extent of receptor-mediated uptake of TGRP was assessed with (a) an anti-apo E monoclonal antibody, which blocks receptor interaction; (b) incubation with heparin; (c) normal vs LDL receptor-negative fibroblasts; and (d) receptor-associated protein (RAP) to determine the potential contribution of LDL receptor-related protein (LRP). Cell surface heparan sulfate proteoglycan- (HSPG-) mediated uptake was examined with or without the addition of heparinase and heparitinase to cell incubation mixtures. At low particle concentrations (250 microg of neutral lipid/mL), most (>/=60%) particle uptake and internalization was via HSPG-mediated pathways. This HSPG pathway did not involve classical lipoprotein receptors, such as LRP or the LDL receptor. These data suggest that in peripheral tissues, such as the arterial wall, apo E may act in TGRP as a ligand for uptake not only via the LDL receptor and LRP pathways but also via HSPG pathways that are receptor-independent. Thus, at physiologic particle concentrations apo E-TGRP can be bound and internalized in certain cells by relatively low affinity but high capacity HSPG-mediated pathways.

No abstract

No abstract

Modification of dietary fatty acid composition results in changes in plasma cholesterol levels in man. We examined the effect of in vitro fatty acid supplementation on low density lipoprotein (LDL) receptor activity in cultured cells and questioned whether changes were related to fatty acid-induced alterations in acyl-CoA: cholesterol acyltransferase (ACAT) activity. Preincubation of cultured cells (i.e. human skin fibroblasts, J774 macrophages, and HepG2 cells) with oleic acid (oleic acid:bovine serum albumin molar ratio 2:1) at 37 degrees C for longer than 2 h resulted in a 1.2- to 1.5-fold increase in LDL cell binding at 4 degrees C and LDL cell degradation at 37 degrees C. Scatchard analysis showed that oleic acid increased LDL receptor number but not LDL affinity (Kd). Fatty acid supplementation of J774 macrophages increased both LDL receptor activity and cholesteryl ester accumulation. The ACAT inhibitor, 58-035, eliminated both effects, and increased ACAT activity preceded stimulation of LDL receptor activity by 1-2 h. Supplementation of macrophages with triolein emulsion particles also increased LDL cell binding and degradation, and addition of cholesterol to the emulsions abolished this effect. Among fatty acids tested, oleate (18:1), arachidonate (20:4), and eicosapentanoate (20:5) demonstrated the greatest effects. We hypothesize that certain fatty acids delivered to cells either in free form, or as triglyceride, first increase cellular ACAT activity, which then causes a decrease in an intracellular free cholesterol pool, signaling a need for increased LDL receptor activity. This mechanism may play a role in the effect of certain dietary fatty acids on LDL metabolism in vivo.