NADPH oxidase (NOX) is a family of oxidases known to produce superoxide and NOX is thought to be involved in neurodegeneration [40]. To determine the role of NOX in TLR3 agonist proinflammatory responses, we investigated the expression of NOX gp91phox, the catalytic subunit of phagocytic oxidase commonly associated with proinflammatory responses. Poly I:C induced NOX gp91phox mRNA 2 to 3 fold (Figure 5A) and NOX gp91phox + IR by manyfold more in cortex and hippocampal dentate gyrus (Figure 5B). Ethanol treatment produced a non-significant trend toward an increase in NOX gp91 mRNA, but did increase gp91phox + IR above control levels to about half of that found with poly I:C alone. Ethanol potentiated the poly I:C TLR3 responses with both NOX gp91phox mRNA and NOX gp91phox + IR increased in cortex and hippocampus (Figure 5). Double antibody studies with cell specific markers and confocal microscopy indicate that ethanol-poly I:C-induced NOX gp91phox + IR is colocalized with neuronal marker (MAP-2) and a microglial marker (Iba1) but there is little colocalization with astrocytic marker (GFAP) (Figure 6). These results indicate that TLR3 activation increases brain NOX gp91phox.
