To explore the mechanism of Huazhuo Xingxue Decoction in inducing cerebral ischemic tolerance(CIT) based on network pharmacology and animal experiment.Methods:The effective components and targets of Huazhuo Xingxue Decoction were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)，Encyclopedia of Traditional Chinese Medicine(ETCM)，Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMAN-TCM)，and literature.The targets of CIT were obtained from GeneCards and Online Mendelian Inheritance in Man(OMIM).Venn diagram was established to identify the targets of Huazhuo Xingxue Decoction in regulating CIT and STRING was employed to build the protein-protein interaction network.Metascape was used for gene ontology(GO) annotation and Encyclopedia of Genes and Genomes(KEGG) for the key targets.The model of cerebral ischemia was established by the Longa thread bolt method in SD rats，which were then intervened with Huazhuo Xingxue Decoction and ischemic preconditioning.Beam walking test(BWT) was employed to evaluate the recovery of motor function，2,3,5-triphenyltetrazolium chloride(TTC) staining to measure the volume of cerebral infarction，and enzyme-linked immunosorbent assay to determine the concentrations of endothelial nitric oxide synthase(eNOS)，neuronal nitric oxide synthase(nNOS)，and inducible nitric oxide synthase(iNOS).Results:Quercetin，kaempferol，phytosterol，luteolin and other compounds in Huazhuo Xingxue Decoction play a synergistic role in inducing ischemic tolerance，involving phosphatidylinositol 3-kinase/protein kinase B(PI3K-AKT)，hypoxia-inducible factor-1(HIF-1)，cyclic adenosine monophosphate(cAMP)，and nuclear factor-κB(NF-κB) signaling pathways and the targets of AKT1，tumor necrosis factor(TNF)，interleukin-6(IL-6)，and tumor protein p53(TP53).Animal experiments showed that the ischemic preconditioning group and Huazhuo Xingxue Decoction group had no differences in the concentration of eNOS，nNOS，and iNOS and the volume of cerebral infarction(P>0.05).The two intervention groups had smaller volume of cerebral infarction and higher concentration of eNOS than the model group(P<0.05).Conclusion:Huazhuo Xingxue Decoction induces CIT in a multi-target and multi-pathway manner and regulates the expression of eNOS.The results of animal experiments verify the results obtained based on network pharmacology，which provides a basis for in-depth study of the CIT induced by Huazhuo Xingxue Decoction.