Background: Luteolin (Lut) is a natural flavonoid with low water solubility. Many studies have revealed that its antitumor effect is more understandable. Objectives: Synthesis of glycyrrhizin-coupled bovine serum albumin-loaded Lut nanoparticles (GL-BSA-Lut-Nps) to progress the water solubility, anticancer effect, liver targeting, cycle arrest, induction apoptosis, and regulating the metabolism of Lut. Materials and Methods: The activity screening of GL-BSA-Lut-Nps on Hepatocellular Carcinoma Bel-7402 cells was spotted by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, cell cycle, and apoptosis were measured by flow cytometry. The solubility of GL-BSA-Lut-Nps was evaluated by ultraviolet spectrophotometer. Fluorescein isothiocyanate was employed to label the drug, and the fluorescence intensity of cells after drug uptake was detected under a fluorescence microscope to sense the targeting of GL-BSA-Lut-Nps to tumor cells. The differences of metabolites between Bel-7402 cells treated with GL-BSA-Lut-Nps and the control group were considered by 1 hydrogen-nuclear magnetic resonance metabolomics. Results: The results presented that the GL-BSA-Lut-Nps prepared by solvent removal method had good anti-tumor activity and water solubility in vitro and the No. 4 Lut nanoparticles (GL-BSA-Lut-Nps-4) screened by the MTT method had the best effect. The IC50 of the GL-BSA-Lut-Nps-4 on the Bel-7402 cell inhibition test was 1.999 ± 0.880 mg/mL. The results of cell cycle and apoptosis displayed that the anticancer effect of the prescription is more palpable. The results of the fluorescence original method are proposed to confirm that the experimentally created GL-BSA-Lut-Nps-4 have a liver targeting effect. The study of metabolomics further clarified the metabolic regulation effect of GL-BSA-Lut-Nps-4 on Bel-7402 cells. Conclusion: It delivers a theoretical basis for the development of new high-efficiency and low-toxicity traditional Chinese medicine preparations for liver cancer.