Nitrosylated high density lipoprotein is recognized by a scavenger receptor in rat liver.
In order to assess the presence of specific recognition sites for high density lipoprotein (HDL) in vivo, HDL was nitrosylated with tetranitromethane and the decay and liver uptake were compared with that of native HDL. The association of intravenously injected nitrosylated HDL (TNM-HDL) with liver was greatly increased as compared to native HDL. Using a cold cell isolation method, it became evident that the liver endothelial cells were responsible for the increased uptake of the modified HDL. The involvement of the endothelial cells in the uptake of TNM-HDL from the circulation could also be demonstrated morphologically by using the fluorescent dye dioctadecyl-tetramethyl-indocarbocyanine perchlorate (Dil) to label HDL. In vitro competition studies with isolated liver endothelial cells indicated that unlabeled modified HDL and acetylated LDL displaced iodine-labeled TNM-HDL, while no competition was seen with LDL and a slight displacement was seen with unlabeled native HDL. Nonlipoprotein competitors of the scavenger receptor such as fucoidin and polyinosinic acid blocked the interaction of TNM-HDL with the liver endothelial cells. Also the degradation of TNM-HDL was blocked by low concentrations of chloroquine. It can be concluded that a scavenger receptor on liver endothelial cells is involved in the clearance of tetranitromethane-modified HDL, which excludes the possibility of using TNM-HDL in vivo to assess the non-receptor-dependent uptake of HDL. The use of nitrosylated HDL in vitro as a low affinity control is limited to cell types that do not possess scavenger receptors, because cell types with scavenger receptors will recognize and internalize TNM-HDL by a high affinity scavenger pathway.
Division of Biopharmaceutics, University of Leiden, The Netherlands.
M F
MF
Kleinherenbrink-Stins
D
D
Schouten
J
J
van der Boom
A
A
Brouwer
D L
DL
Knook
T J
TJ
van Berkel
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004727:Endothelium / Q000378:metabolism; D005456:Fluorescent Dyes; D007728:Kupffer Cells / Q000378:metabolism; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013774:Tetranitromethane / Q000378:metabolism; D013997:Time Factors
D005456:Fluorescent Dyes; D008075:Lipoproteins, HDL; D013774:Tetranitromethane