10 000 HEK-293Tcells seeded in 96 well plates were transfected 24 h later by using 0.2 µl of jetPRIME

OBJECTIVE: Macrophages, converted to lipid-loaded foam cells, accumulate in atherosclerotic lesions. Macrophage lipid metabolism is transcriptionally regulated by peroxisome proliferator-activated receptor gamma (PPARgamma), and its target gene fatty acid binding protein 4 (FABP4) accelerates the progression of atherosclerosis in mouse models. Since expression of PPARgamma and FABP4 is increased upon interleukin-4 (IL-4)-induced macrophage polarization, we aimed to investigate the role of FABP4 in human IL-4-polarized macrophages. METHODS AND RESULTS: We investigated the impact of FABP4 on PPARgamma-dependent gene expression in primary human monocytes differentiated to macrophages in the presence of IL-4. IL-4 increased PPARgamma and its target genes lipoprotein lipase (LPL) and FABP4 compared to non-polarized or LPS/interferon gamma-stimulated macrophages. LPL expression correlated with increased very low density lipoprotein (VLDL)-induced triglyceride accumulation in IL-4-polarized macrophages, which was sensitive to inhibition of lipolysis or PPARgamma antagonism. Inhibition of FABP4 during differentiation using chemical inhibitors BMS309403 and HTS01037 or FABP4 siRNA decreased the expression of FABP4 and LPL, and reduced lipid accumulation in macrophages treated with VLDL. FABP4 or LPL inhibition also reduced the expression of inflammatory mediators chemokine (C-C motif) ligand 2 (CCL2) and IL-1beta in response to VLDL in IL-4-polarized macrophages. PPARgamma luciferase reporter assays confirmed that FABP4 supports fatty acid-induced PPARgamma activation. CONCLUSION: Our findings suggest that IL-4 induces a lipid-accumulating macrophage phenotype by activating PPARgamma and subsequent LPL expression. Inhibition of FABP4 decreases VLDL-induced foam cell formation, indicating that anti-atherosclerotic effects achieved by FABP4 inhibition in mouse models may be feasible in the human system as well.