Transfer of [ 3H]cholesterol between lipid vesicles and rat arterial smooth muscle cells in vitro

Abstract

Unesterified [ 3H]cholesterol is rapidly transferred between cholesterol-phosphatidylcholine vesicles and rat arterial smooth muscle cells in vitro. Exchange rate is influenced by the vesicle/cell ratio in a saturable way. The maximal transfer of cholesterol, which is 3.76 μug per mg cell protein during 4 h, is achieved with a vesicle/cell ratio of 3.4 · 10 7. Bovine serum albumin enhances the exchange by a factor of 4.5 compared to a protein-free system. The activation energy for the process is + 38.5 kJ · mol −1 with vesicles of 1:1 mole ratio of cholesterol to phosphatidylcholine (C/P). A fraction of the incorporated free [ 3H]cholesterol is esterified within 4 h with donor vesicles of over 1:1 C/P. When cells were incubated with vesicles of low C/P mole ratio (1:2) a fraction of the incorporated free [ 3H]cholesterol was esterified within 16 h. Our results are compatible with the aqueous diffusion mechanism of cholesterol exchange. Furthermore, we suggest that, in rat smooth muscle cells, the cell membrane cholesterol pool is not metabolically isolated from internal cholesterol pools, at least as judged by the ability of the cells to esterify incorporated free cholesterol.