Figure 2

Anti-inflammatory effects of diazoxide pre-treatment in microglial cell cultures stimulated with LPS and IFNγ. Nitrite accumulation (A), and TNF-α (B) and IL-6 release (C) in control (unstimulated cells), DZX (unstimulated cells pretreated with 100 μM diazoxide), diazoxide (DZX) and LPS/IFNγ + Diazoxide (10 μM to 100 μM) normalized for LPS/IFNγ untreated cells. Quantification of iNOS (D) and COX-2 (E) protein expression in control, LPS/IFNγ untreated cells and 100 μM diazoxide pre-treated LPS/IFNγ cells. Protein expression was measured by western blot and data normalized with β-actin. Images showing representative immunoblotting (F). Percentage of phagocytic cells quantificated by fluorescent microspheres incorporation of control, LPS/IFNγ untreated cells and 100 μM diazoxide pre-treated LPS/IFNγ cells (G). One representative phagocytosis experiment is shown (H,left). Phagocytosis of microspheres is represented by the peaks at the high fluorescence levels (H,right) Control: unstimulated cells; DZX: unstimulated cells pretreated with 100 μM diazoxide; L+I: cells stimulated with LPS and IFNγ; L+I+DZX: L+I-stimulated cells pretreated with diazoxide. Results are shown as mean ± SEM of three to five independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs L+I for A-E and vs control for G.