Conformational changes in oxidized LDL recognized by mouse peritoneal macrophages

Abstract

Mouse peritoneal macrophages have been considered to recognize and take up oxidized LDL by a scavenger receptor. However, it is still unknown what conformational changes in oxidized LDL contribute to recognition by the macrophage scavenger receptor. In the present study, it was shown that the amount of oxidized LDL taken up by macrophages correlated well with the fluorescence intensity formed in oxidized LDL. The autofluorescent products generated in oxidized LDL were characterized by Ex:365 nm Em:430 nm, and the intensity of the fluorescence was reduced at base pH, and restored by adjusting the pH to neutral. The characteristics of the fluorescent products indicate that a Schiff base structure was formed in oxidized LDL. Oxidized LDLs were fractionated into native size and aggregated large particles with HPLC by monitoring fluorescence. It was demonstrated that macrophages ingest selectively or preferentially aggregated oxidized LDL, but not native size oxidized LDL. The incorporation of aggregated oxidized LDL was remarkably suppressed by heparin and cytochalasin B. These results suggest that mouse peritoneal macrophages recognize the conformational changes in oxidized LDL related to the formation of a Schiff base structure with increasing autofluorescence, and ingest selectively aggregated large particles in oxidized LDL in a phagocytic process.