To explore the active components of Chinese medicines(TCM) in regulating glycolysis and their medication rules.Methods:Relevant targets associated with glycolysis were obtained from the GeneCards database.The DAVID database was used to analyze the biological processes and signaling pathways involving these targets.Target information was then imported into the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform(TCMSP) to identify corresponding active components and Chinese medicines，and a “target-active component-Chinese medicine” network was constructed.The properties，flavors，meridian tropism，and effects of these Chinese medicines were statistically analyzed to determine medication rules for regulating glycolysis.Finally，cell experiments were conducted to verify the effects of Salviae Miltiorrhizae Radix et Rhizoma and Astragali Radix on the expression of glycolysis-related proteins in gastric cancer precursor cells(MC).Results:A total of 24 glycolysis-related targets were identified，matching 238 active components and 337 Chinese medicines.Key effective components that may intervene in glycolysis include quercetin，kaempferol，and luteolin，while major Chinese medicines identified include Glycyrrhizae Radix et Rhizoma，Uncariae Ramulus cum Uncis，and Salviae Miltiorrhizae Radix et Rhizoma.The analysis of medication rules revealed that the medicinal properties were predominantly cold or warm; the flavors were mainly bitter or sweet; these Chinese medicines primarily acted on the liver and lung meridians; the main effects were clearing heat and tonifying deficiency.Experimental results indicated that both Salviae Miltiorrhizae Radix et Rhizoma and Astragali Radix significantly reduced the expression of glycolysis-related proteins，including hypoxia-inducible factor-1α(HIF-1α)，glucose transporter 1(GLUT1)，hexokinase 2(HK2)，pyruvate kinase M2(PKM2)，and lactate dehydrogenase A(LDHA) in MC cells.Conclusion:This study utilized bioinformatics method to identify active components and medication rules of Chinese medicines in regulating glycolysis，aiming to provide insights for formulating treatments for glycolysis-related diseases，as well as reference for the development of new Chinese medicines and mechanistic studies.