Acute effects of oxidized low density lipoprotein on metabolic responses in macrophages.
The immediate effects of oxidized low density lipoprotein (OxLDL) on the metabolic activity of cultured macrophages (RAW 264.7) were studied using a microphysiometer. Administration of OxLDL acutely induced a concentration-dependent increase in metabolic activity, with an EC50 of 16 +/- 3 microg/ml OxLDL and a maximal effect of 35% +/- 4% (mean +/- SEM; n=5). A biphasic response was measured after administration of 75 or 100 microg/ml OxLDL consisting of an initial sharp increase, followed by the induction of a long-lasting hypoactivity of 80% of the control value. Incubation of cells with polyinosinic acid (polyI; 100 microg/ml) for 30 min prior to OxLDL administration could completely block the effect of 25 microg/ml OxLDL. In addition, polyI acted as a full antagonist on the decrease of the biphasic response of cells generated by 75 and 100 microg/ml OxLDL. Macrophages used in this study possessed a specific binding site for OxLDL, with a dissociation constant (KD) of 9 +/- 2 microg/ml and a maximal binding of 610 +/- 32 ng 125I-OxLDL/mg cell protein. Binding of 125I-OxLDL to macrophages could be completely competed for by unlabeled OxLDL, by polyI for 58%, and by AcLDL for 46%. In conclusion, OxLDL can acutely activate the metabolic state of macrophages by a receptor-mediated process in a concentration-dependent fashion, which could be antagonized by polyI. Metabolic responses to OxLDL may underlie the changes observed in macrophages in the early atherosclerotic plaque.
De Vries
H E
HE
Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, University of Leiden, The Netherlands. e.vries@lacdr.LeidenUniv.nl
Ronken
E
E
Reinders
J H
JH
Buchner
B
B
Van Berkel
T J
TJ
Kuiper
J
J
D000818:Animals; D002478:Cells, Cultured; D006863:Hydrogen-Ion Concentration; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008244:Lysophosphatidylcholines / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism*; D008565:Membrane Proteins; D051379:Mice; D010084:Oxidation-Reduction; D011485:Protein Binding; D011971:Receptors, Immunologic / Q000378:metabolism; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; D051122:Scavenger Receptors, Class B
D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines; D008565:Membrane Proteins; D011971:Receptors, Immunologic; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B; C099252:oxidized low density lipoprotein