Activation of NADPH oxidase (NOX) produces superoxide in brain and superoxide formation was assessed in the present experiment by histochemistry using in situ visualization of reactive oxygen species, for example, O2- and O2--derived oxidant production of ethidium from hydroethidine [32,33]. In vehicle-treated mice, there was little to no detection of O2- and O2--derived oxidant production of ethidium (Figure 7). In mice that received either ethanol or poly I:C treatment, there was a significant increase in the production of O2- and O2--derived oxidants 27 hours after the last dose of ethanol or three hours after poly I:C administration. Pretreatment with ethanol increased poly I:C production of O2- and O2--derived oxidants 3 to 4 fold above poly I:C alone and more than 10 fold over controls (Figure 7). These findings indicate that ethanol increases expression of NOX gp91phox and the formation of reactive oxygen species by TLR3 agonist.
