Cell culture
For macrophage differentiation, monocytes were cultured at 37°C with 5% CO2 in 24-well plates (Costar) for 6 days in supplemented RPMI medium (10% fetal bovine serum, 10nM glutamine, 100 μg/ml streptomycin, 100 U/ml penicillin) and 50 ng/ml GM-CSF for M1 macrophages, then incubated for 24 hours with IFN-γ (100 ng/ml) and LPS (100 ng/ml). For MMe and M2 macrophages, monocytes were incubated for 6 days with M-CSF (50 ng/ml), and then incubated for 24 hours with 15mM glucose (Sigma Aldrich), 10nM insulin (Sigma Aldrich) and 0.4mM palmitate (Sigma Aldrich) to polarize to MMe, or with IL-4 (20 ng/ml) to obtain M2 macrophages. Cells were harvested using EDTA and further evaluation by flow cytometry was performed. Culture media were collected for cytokine measures.