Abstract
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 ...Continue Reading
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