Reverse cholesterol transport is regulated by varying fatty acyl chain saturation and sphingomyelin content in reconstituted high-density lipoproteins.

Because phospholipid composition of high-density lipoprotein (HDL) plays a vital role in its reverse cholesterol transport (RCT) function, we studied RCT in vitro (uptake and efflux) with reconstituted HDLs (rHDLs) containing phosphatidylcholine (PC) with fatty acids of increasing saturation levels (stearic, oleic, linoleic, linolenic) and without or with sphingomyelin (SM). Uptake significantly increased from basal value when the PC component included up to 50 mol % of oleic or linolenic acid, but did not change with linoleic acid. Increasing oleic and linoleic acids to 100 mol % significantly decreased uptake, but increasing linolenic acid to the same value did not affect it. Sphingomyelin in rHDL significantly decreased uptake, but only with PC-containing unsaturated fatty acids, and not with saturated fatty acid. Efflux was not affected in a dose-dependent manner when oleic or linoleic acid content was increased, but was significantly increased with levels of linolenic acid up to 25 mol % in PC, and was dramatically lowered with higher levels. Sphingomyelin in rHDL (PC/SM, 20:80, mol/mol) significantly increased efflux only with oleic or linoleic acid-containing rHDLs, compared with efflux without SM. In conclusion, enrichment of PC component up to 25 mol % as linolenic acid has a beneficial effect on RCT, whereas a higher percentage of it or other unsaturated fatty acids seems to be detrimental. In addition, high SM content decreases uptake with rHDL-containing unsaturated fatty acids, whereas it increases efflux for rHDL-containing oleic or linoleic acid. These results show for the first time the importance of SM in RCT in a well-defined in vitro system.