Japanese encephalitis virus (JEV) infection is commonly associated with inflammatory reaction and neurological disease that occurs in the infected animals. Reactive oxygen species have been implicated as a critical mediator for inflammation and diseases. The present study investigated the change of redox potential in glial cells following JEV infection. JEV infection induced the generation of superoxide anion and nitric oxide in rat cortical glial cells. Manganese superoxide dismutase, but not copper/zinc superoxide dismutase was activated by JEV infection, and this activation was blocked by pyrrolidine dithiocarbamate. In addition, the increased superoxide dismutase activity was also apparent in JEV acutely, or persistently infected continuous cell lines. These results suggest that cellular factors regulating oxidative pathway might play roles in responding to JEV infection.