This study investigates the active ingredients and mechanism of action of Gynostemma pentaphyllum in the treatment of chronic fatigue syndrome by using network pharmacology and animal model validation methods.The active components and target of G. pentaphyllum were extracted from TCMSP,TCMID,BATMAN-TCM and SWISS-TARGET databases;Disease targets were obtained from OMIM and GeneCards databases;select the intersection targets after mapping drug targets to disease targets and draw the active ingredient-shared target network;The PPI network was constructed by String database and Cytoscape3.7.1;GO and KEGG enrichment analysis of intersection targets by R language;Autodock Vina was used to conduct molecular docking between core components and key targets;Establishment of a mouse model of chronic fatigue syndrome,and the therapeutic effect of G. pentaphyllum on chronic fatigue syndrome was verified by animal test.The results finally obtained 17 active ingredients of G. pentaphyllum,corresponding to 583 targets,the number of targets of chronic fatigue syndrome is 2 692,the total number of target screening results are 266,the number of core targets is 96.The enrichment pathways mainly include P13K-AKT,lipid and atherosclerotic,fluid shear stress and atherosclerotic signaling pathways.Molecular docking results showed that the core components of G. pentaphyllum had good binding force with the key target of CFS.Animal experiments showed that,compared with the model group, G. pentaphyllum could prolong the exercise time of mice and significantly reduce the serum urinary nitrogen level and lactate dehydrogenase activity of mice.Western blot showed that G. pentaphyllum could significantly up-regulate the expression of PI3K and AKT proteins in the muscle tissue of chronic fatigue mice,and significantly down-regulate the expression of P-PI3K,P-AKT and STAT3 proteins.In summary,the present study preliminary suggests that  G. pentaphyllum may play a role in the treatment of chronic fatigue syndrome by regulating the core targets such as STAT3,TP53,SRC,AKT1,HSP90AA1 and the PI3K/AKT signaling pathway.