To analyze the potential mechanism of Jingxin Zhidong Formula(JXZDF) in treating tic disorder(TD) in children through network pharmacology and verify the prediction results in vitro.Methods:The active components of JXZDF were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and the Encyclopedia of Traditional Chinese Medicine(ETCM)，and the targets of the components from SwissTargetPrediction.The TD disease targets were obtained from GeneCards，Online Mendelian Inheritance in Man(OMIM)，Therapeutic Target Database(TTD)，Pharmacogenetics and Pharmacogenomics Knowledge Base(PharmGkb)，and DisGeNET.The common targets shared by the active components of JXZDF and TD were taken as the potential target of JXZDF in treating TD.The common targets were imported into STRING to build the protein-protein interaction(PPI) network.The “Chinese medicine-active component-disease target” network was built in Cytoscape，and the core target was screened by the plug-in of CytoNCA.Gene ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment were carried out for the core targets.Furthermore，the activated microglia were treated with JXZDF.Western blotting was then employed to verify the targets of JXZDF in intervening in microglia.Results:The network pharmacology predicted that the main active components of JXZDF were quercetin，stigmasterol，hispidulin，β-sitosterol，kaempferol，etc.，and JXZDF may treat TD via the PI3K/AKT signaling pathway.The cell experiment showed that JXZDF down-regulated the expression of p-PI3K/PI3K and p-AKT/AKT in activated microglia(P<0.05).The combination of JXZDF with PI3K agonist 740Y-P reversed such down-regulation(P<0.05).Conclusion:This study predicted the potential signaling pathway of JXZDF in treating TD and confirmed that JXZDF interfered with the function of microglia by down-regulating the PI3K/AKT signaling pathway，providing evidence for further study of JXZDF in treating TD.