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.