Metabolically activated macrophages from patients and controls produce equal amounts of cytokines.
To evaluate the MMe function, we recollected supernatants from the macrophage cultures treated with metabolic stimuli in order to determine concentration of different cytokines. First, we compared the cytokines levels produced by pro-inflammatory macrophages M1 and MMe. In the control group, we found higher levels of IL-12 (p= 0.0156, 0.97%, 0.33%-1.67% vs 2.07%, 1.71%-2.15%, median and IQR; Fig 6A) in the MMe supernatants, while IL-1β, IL-6, IL-8, IL-10 and TNFα were equally produced by MMe and M1 macrophages (data not shown). Interestingly, in dyslipidemia patients (Fig 6B), we found a higher concentration of IL-10 in the MMe supernatants (p= 0.0391, 4.21%, 2.4%-4.61% vs 56.33%, 2.98%-104%, median and IQR) and higher IL-6 levels in M1 supernatants (p= 0.0156, 390.8%, 1726%-55.84% vs 273.5%, 570.8%-31.3%, median and IQR). Since it is known that when culturing monocytes with metabolic stimuli there is also differentiation not only towards MMe, but also towards M1 and M2 macrophages, we evaluated the intracellular expression of IL-10 by flow cytometry in MMe to corroborate that these cells, in fact, produce IL-10 (85.8%, 82.5%-86.3%, median and IQR; Fig 6B). When we evaluated cytokine concentrations, produced by MMe from healthy and dyslipidemia subjects, we did not observe differences between groups (NS, Fig 6C). Additionally, we did not find differences among the cytokine concentrations in serum samples from healthy and dyslipidemia subjects, with the exception of IL-12 (p= 0.0003, 0.43%, 0.52%-0.12% vs 3.12%, 4.05%-2.63%, median and IQR, Fig 6D), that was higher in the group with dyslipidemia.