Redox effector factor (Ref-1) is a sensitive marker for oxidative cellular injury. The aim of this study was to investigate the effects of acupuncture on hippocampal Ref-1 expression in cerebral multi-infarction rats. The rats with reference memory impairment were randomly allocated to three groups: impaired group, acupuncture group and placebo acupuncture group. Moreover, normal group and sham-operated group were set as control groups. Morris water maze test showed that cerebral multi-infarction rats did not present significant changes in spatial working memory performance. Further investigation by immunohistochemistry revealed that acupunctural treatment significantly increased the expression of Ref-1 in the hippocampus of the impaired rats. These findings suggested that the spatial working memory was unaffected in the cerebral multi-infarction rats although spatial reference memory deficits were detected in our previous study; in addition, acupuncture could increase the Ref-1 expression, consequently exerting the anti-oxidant effects.