Abstract: Objective: To investigate the neuroprotective effect of sanguinarine on ischemic stroke rats by regulating sphingosine kinase 1(SphK1)/sphingosine-1-phosphate(S1P) signaling pathway. Methods: Seventy-five SD rats were randomly grouped into sham surgery group, model group, sanguinarine group, PF543 group(SphK1 inhibitor), and sanguinarine+PMA(SphK1 activator) group, with 15 rats in each group. Except the sham surgery group, the rat model of ischemic cerebral infarction was established in other groups. After modeling, neural function of rats in each group was evaluated, and pathological changes of hippocampal morphology was observed by hematoxylin-eosin(HE) staining. The water content of brain tissue and the level of malondialdehyde(MDA), superoxide dismutase(SOD) and catalase(CAT) in brain tissue were detected. The cerebral infarction area was detected by TTC method, the neuronal apoptosis rate was detected by TUNEL method, and the protein expression of Bax, Bcl-2 and SphK1 were detected by western blotting. Results: In the model group, the hippocampal neurons of rats were seriously damaged, the neurological deficit score, brain tissue water content, the level of MDA,the expression levels of Bax and SphK1 proteins, cerebral infarction area, and the neuronal apoptosis rate were obviously increased, and the activities of SOD, CAT, and the expression level of Bcl-2 protein in brain tissue were obviously reduced(P＜0.05); the neurological deficit score, brain tissue water content, the level of MDA,the expression levels of Bax and SphK1 proteins, cerebral infarction area, neuronal apoptosis rate were obviously reduced, and the activities of SOD, CAT,and the expression level of Bcl-2 protein in brain tissue were obviously increased in the sanguinarine and PF543 groups(P＜0.05). SphK1 activator PMA partially reversed the protective effect of sanguinarine on rats. Conclusion: Sanguinarine may have a neuroprotective effect on ischemic stroke rats by inhibiting oxidative stress and neuronal apoptosis and blocking the SphK1/S1P signaling pathway.