The study explores the molecular mechanism of nuciferine in inhibiting the proliferation of human intrahepatic cholangiocarcinoma cell HuCCT1 by network pharmacology.The MTT colorimetric and clone formation assay was used to measure the proliferation activity of HuCCT1 cells.After nuciferine treatment,the glycolytic biochemical indexes were detected by colorimetric and spectrophotometric assay.The potential targets and pathways of nuciferine in intrahepatic cholangiocarcinoma were predicted by constructing a network and the enrichment analysis results.And further investigation of nuciferine inhibiting HuCCT1 cell proliferation is being explored based on it.It is shown that nuciferine improved HuCCT1 proliferation in a time-and dose-dependent manner and significantly reduced cellular glucose consumption and lactate production.Network pharmacological analysis indicated that nuciferine affected intrahepatic cholangiocarcinoma growth and motility mainly through PI3K/ Akt,MAPK,and HIF-1 signaling pathways.Data of polymerase chain reaction (PCR) and Western blot demonstrated that nuciferine was able to downregulate the mRNA and protein expression of hypoxia-inducible factor-1α (HIF-1α),hexokinase 2 (HK2),pyruvate kinase 2 (PKM2),as well as protein expression of p-AktThr308,p-mTOR and p-4EBP1.In conclusion,nuciferine may inhibit the proliferation of HuCCT1 cells through the adjustment of glycolytic activity regulated by the Akt/mTOR/4EBP1 signaling cascade.