Human macrophage-mediated oxidation of low-density lipoprotein is delayed and independent of superoxide production.
There is growing evidence that oxidatively modified low-density lipoprotein (LDL) accumulates in the atherosclerotic intima of arteries. Cells present in the intima (including the monocyte/macrophage) are capable of oxidizing LDL in vitro, but the mechanisms by which this occurs are unknown. Several reports have claimed a crucial role for superoxide as a cell-derived radical species capable of enhancing the rate of LDL oxidation. We have used a sensitive h.p.l.c. system with chemiluminescence detection to measure LDL cholesteryl ester hydroperoxides at early stages of LDL oxidation. During the initial stages of LDL oxidation, there is at least a 2 h delay before human monocyte-derived macrophages enhance this process. Stimulation of these cells to produce large fluxes of superoxide does not increase the rate of LDL oxidation or decrease the delay of its onset. Prior exposure of LDL to a high flux of superoxide does not increase its susceptibility to oxidation by human monocyte-derived macrophages. We also show that the thiobarbituric acid-reactive substances (TBARS) assay does not always correlate with more direct methods of assessing LDL oxidation and confirm recent reports that superoxide dismutase only partially inhibits cell-mediated LDL oxidation. We conclude that superoxide does not play a major role in human monocyte-derived macrophage-mediated LDL oxidation under the conditions that we describe.
Garner
B
B
Cell Biology Group, Heart Research Institute, Sydney, N.S.W., Australia.
Dean
R T
RT
Jessup
W
W
D002788:Cholesterol Esters / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D015227:Lipid Peroxidation; D008077:Lipoproteins, LDL / Q000097:blood*; D008163:Luminescent Measurements; D008264:Macrophages / Q000378:metabolism*; D010084:Oxidation-Reduction; D016897:Respiratory Burst / Q000187:drug effects; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000097:blood*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D002788:Cholesterol Esters; D008077:Lipoproteins, LDL; D017392:Thiobarbituric Acid Reactive Substances; D013481:Superoxides; D013482:Superoxide Dismutase; D013755:Tetradecanoylphorbol Acetate