Cholesterol accumulation in aortic smooth muscle cells exposed to low density lipoproteins. Contribution of free cholesterol transfer.
Incubation of cultured arterial smooth muscle cells with large concentrations of low density lipoproteins (LDL) resulted in a net increase in cell cholesterol and cholesteryl ester mass that was dependent on LDL concentration and time of incubation. Use of an inhibitor of acyl-CoA:cholesterol acyl-transferase (ACAT) reduced the accumulation of cholesteryl ester mass by 40% (range 25% to 50%), suggesting that a significant proportion of the cholesteryl ester mass that accumulated from LDL did so without being hydrolyzed and re-esterified. Quiescent arterial smooth muscle cells exposed for 48 hours to 0.5 mg/ml of 125I-LDL accumulated 115 nmol total sterol/mg cell protein. However, these cells took up and degraded only 21 micrograms of 125I-LDL protein, which contains 64 nmol total cholesterol. Hence, only about 60% of the increase in cell-associated cholesterol mass was accounted for by LDL particle uptake and degradation. Further, when cells were incubated with 3H cholesteryl linoleyl ether-labeled LDL, the net increase of total cell cholesterol was 81 nmol/mg cell protein. However, only 49 nmol of total cholesterol was taken up by LDL particle uptake, as calculated from the uptake of the 3H cholesteryl linoleyl ether tracer. It thus appears that about 40% of the accumulated cholesterol mass was derived independent of LDL particle uptake, suggesting the possibility of transfer of free cholesterol from the surface of LDL to the cell surface. The occurrence of cholesterol surface transfer was independently verified by the measurement of the uptake and cellular distribution of LDL-derived free 3H-cholesterol. A substantial fraction of the accumulated cell cholesterol mass (approximately 40%) was derived from surface transfer of LDL free cholesterol.(ABSTRACT TRUNCATED AT 250 WORDS)
Slotte
J P
JP
Department of Medicine, University of Washington, Seattle 98195.
Chait
A
A
Bierman
E L
EL
D000818:Animals; D001011:Aorta; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002784:Cholesterol / Q000031:analogs & derivatives / Q000378:metabolism* / Q000502:physiology; D002788:Cholesterol Esters / Q000378:metabolism; D002789:Cholesterol Oxidase; D003593:Cytoplasm / Q000378:metabolism; D006801:Humans; D008077:Lipoproteins, LDL / Q000502:physiology*; D008254:Macaca nemestrina; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*
D002788:Cholesterol Esters; D008077:Lipoproteins, LDL; C027575:cholesteryl linoleyl ether; D002784:Cholesterol; D002789:Cholesterol Oxidase