〔Abstract〕 Objective To explore the regulatory mechanism of ginsenoside Rb1 on focal cerebral ischemia-reperfusion injury(CIRI). Methods A total of 60 C57/BL mice were randomly divided into 6 groups(n=10): sham-operated group, CIRI model group, ginsenoside Rb1 low-, medium-, and high-dose group and nimodipine(positive control) group. The surgical method was used to construct the focal CIRI mouse model. The neurological function scores and behavioral tests were performed, and Nissl staining was utilized to detect the number of nissl bodies in the hippocampus. The effect of ginsenoside Rb1 on the molecule expression of the Wnt signaling pathway in the hippocampus was detected by qPCR, Western blot and immunohistochemistry assays. The regulatory mechanism of ginsenoside Rb1 was investigated through molecular docking and co-precipitation assays. Results Compared with the CIRI model group, the addition of ginsenoside Rb1 reduced the neurological function scores of mice(P<0.05), shortened the time passing the balance beam(P<0.05), but increased the time entering the correct arm(P<0.05) and the swinging time and climbing time of mice(P<0.05), indicating that ginsenoside Rb1 could effectively resume the function of the nervous system in mice and improve the behavioral ability of model mice. After ginsenoside Rb1 treatment, axis inhibition protein 2(Axin2) and glycogen synthase kinase-3β(GSK-3β) in the hippocampus decreased, whereas the expression of Wnt3a, Wnt1 and β-catenin increased. Conclusion The ginsenoside Rb1 can improve neurological function of the CIRI mouse model and increase the number of Nissl bodies in the hippocampus, which is correlated with the activation of the Wnt signaling pathway, and it may be neuroprotective against focal CIRI during stroke treatment.