Oxidation-induced aggregation of rabbit low-density lipoprotein by azo initiator

Oxidation of low-density lipoprotein (LDL) has been considered as an important step in the early pathogenesis of atherosclerosis. We investigated the oxidative modification of LDL by a water-soluble azo-initiator AAPH (2,2,-azo-bis(2-amidinopropane).2HCl) and analyzed the uptake of AAPH-oxidized LDL with mouse peritoneal macrophages. Oxidative modification of LDL by AAPH was similar to the modification induced by copper in regard to the degree of oxidation and formation of aggregated LDL. The aggregated oxidized (AO-) LDL was fractionated by gel permeation chromatography and compared with the monomeric oxidized (MO-) LDL to make clear their characterization. The results of binding, cell association, and degradation with macrophages indicated that both AO- and MO-LDL were bound and endocytosed by macrophages. The cross competition experiment showed that nonreciprocal competition existed among MO-LDL, AO-LDL, and monomeric acetylated (MAc-) LDL. By the sterol accumulation experiment in macrophages with the various types of modified LDL, the cellular sterol accumulation was shown as the following order, AO-LDL > MAc-LDL > MO-LDL. These results indicated that the oxidation by AAPH can induce the aggregation of LDL and that the AO-LDL contribute to lipid accumulation into macrophages more than the MO-LDL.