Figure 5

AICAR inhibits TNF-α-, and/or IL-1β- and/or Aβ- (25–35) stimulated iNOS expression and nitric oxide release in astrocytic cellcultures. Cells were pre-incubated with 1 mM AICAR prior to stimulation. Fig. A shows nitric oxide released into the medium upon treatment with cytokines +/- Aβ (25–35) with relevant controls. Figure B shows nitric oxide released into the medium and corresponding western-immunoblot for iNOS, COX-2, and MnSOD, after stimulation with cytokines (TNF-α + IL-1β ± Aβ peptide), either in the presence or absence of AICAR in concentrations used in figure A. Figure C shows a dose-dependent inhibition of nitric oxide production and expression of iNOS, COX-2 and MnSOD proteins, on stimulation with cytokines and Aβ and after pre-incubation with increasing amounts of AICAR, as shown. The increase in p-AMPK protein band (Thr-172) indicates activation of AMPK with increasing concentrations of AICAR.