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Abstract
Oxidation of LDL is now widely accepted to be involved in atherogenesis. The aim of this study was to examine the effect of BO-653, a strong radical scavenger and antioxidant, on oxidation of LDL by human macrophages in vitro. Fifty μg/ml LDL protein was incubated with macrophages in Ham's F10 medium, supplemented with additional Fe2+, for up to 48h. Then the medium was analysed by LDL agarose gel electrophoresis, the thiobarbituric acid assay and gas chromatography. In the absence of added exogenous antioxidants, after 24 h LDL oxidation produced 30.48 nmoles MDA equivalents/mg LDL protein and a relative electrophoretic mobility of 4.74. Linoleic acid (18:2), arachidonic acid (20:4) and cholesterol were depleted and 7β-hydroxycholesterol was generated. BO-653 completely inhibited this cell-mediated oxidation of LDL in concentrations as low as 5 μM, being more effective than either α-tocopherol or probucol, which completely inhibited oxidation at 200 and 80 μM and only partially at 80 and 8 μM, respectively. This inhibition of cell-mediated LDL oxidation was not due to toxicity, as α-tocopherol, probucol and BO-653 were not toxic for the macrophages at the concentrations tested. Eighty μM α-tocopherol, 8 μM probucol and 5 μM BO-653 significantly reduced the toxicity to the oxidising culture caused by LDL oxidation. The results show that in this system BO-653 is a more effective antioxidant than α-tocopherol or probucol.