Ac-hE-18A-NH2, a novel dual-domain apolipoprotein mimetic peptide, inhibits apoptosis in macrophages by promoting cholesterol efflux

Abstract

A novel synthetic dual-domain apolipoprotein (apo)-mimetic peptide, Ac-hE-18A-NH2, has been proposed to possess several apo A-I-and apo E-mimetic properties. This study investigated the protective effect of this peptide on oxidized low-density lipoprotein (ox-LDL)-induced apoptosis in RAW264.7 cells. For this purpose, RAW264.7 cells were exposed to 50 μg/ml ox-LDL for 48 h, and then incubated with the peptide Ac-hE-18A-NH2 at various concentrations. Apoptosis was detected using annexin V-fluorescein isothiocyanate staining and flow cytometric analysis. The study revealed that the peptide Ac-hE-18A-NH2 (1, 10 and 50 μg/ml) inhibited ox-LDL-mediated apoptosis, and this was accompanied by an increased rate of intracellular cholesterol efflux, and decreased total cholesterol levels in the cells in a concentration-dependent manner. The peptide also decreased caspase-3 activity and increased B-cell lymphoma 2 protein (Bcl-2) expression in macrophages in a dose-dependent manner. Moreover, blockage of cholesterol efflux by brefeldin A decreased the protective effect of Ac-hE-18A-NH2 against ox-LDL induced apoptosis, while increasing the cholesterol efflux by β-cyclodextrin administration led to a marked decrease in the rate of apoptosis of the cells. These findings demonstrate that the apo-mimetic peptide Ac-hE-18A-NH2 exerts a protective effect against apoptosis by reducing the accumulation of cholesterol.