Fig. 4

LL-37, HBD2 and IL-10 gene expression and NO production by monocytes infected from T2D patients and healthy volunteers. A total of 0.5 × 10⁵ monocytes/well were placed in a culture plate and infected with M. tuberculosis at MOI 10 for 1 h, extracellular M. tuberculosis was eliminated by washing, and the monocytes were incubated for 1 h (D0), 24 h (D1) and 72 h (D3). The supernatants were harvested, and the cells lysed for RNA extraction and cDNA synthesis. LL-37 and IL-10 gene expression was assessed using qPCR and HBD2 was analysed by PCR. In the supernatants, the nitrite (−NO₂) concentration was determined by Griess reagent and reported as μM. a The dot graph shows the LL-37 gene expression in T2D patients, n = 11(closed circles) and Healthy volunteers, n = 11(open circles) at D3. p > 0.5, T2D vs. Healthy (Mann-Whitney U test). b The dot graph shows the HBD2 gene expression in T2D patients, n = 11 and Healthy volunteers, n = 11 in uninfected (UN) and infected (M.tb) monocytes at D3. p > 0.05, T2D vs. Healthy (Mann-Whitney U test). c The image shows representatives amplifications of HBD-2 and HPRT fragments used as housekeeping gene in UN and M.tb monocytes from T2D (n = 3) and Healthy volunteers (n = 3) at D3. d The dot graph shows the IL-10 gene expression in T2D patients, n = 11 and Healthy volunteers, n = 11 at D3. p > 0.05, T2D vs. Healthy (Mann-Whitney U test). e The dot graph shows the NO production by monocytes from T2D patients, n = 30 and Healthy volunteers, n = 19 at D3. p > 0.05, T2D vs. Healthy (Mann-Whitney U test). f NO production by monocytes from T2D patients with levels of 25OHD₃ < 50 nM, n = 22, >50 nM, n = 8 and Healthy volunteers, n = 19 at D0 and D3. *p ≤ 0.05, T2D, 25OHD₃ < 50 nM and ≥50 nM and Healthy at D0 vs. D3 (Wilcoxon test); *p ≤ 0.05, T2D, 25OHD₃ < 50 nM vs. Healthy at D3 (Mann-Whitney U test). In all graphics the median is represented by horizontal line