Aim To investigate the effect of ezetimibe on hepatic low density lipoprotein cholesterol (LDLC) uptake and its mechanism. Methods The hyperlipidemia model mice had intragastric administration of 5 mg/(kg·d) ezetimibe. After four months, the contents of total cholesterol (TC), LDLC and high density lipoprotein cholesterol (HDLC) were measured in the blood of mice, and liver tissue morphology was observed by HE staining. HepG2 cells were incubated with ezetimibe (30 μmol/L) and low density lipoprotein (25 mg/L) for 24 hours. The distribution of lipid droplets in cells was observed by oil red O staining. Enzymatic method was used to detect the contents of intracellular TC, free cholesterol (FC) and cholesteryl ester (CE). The uptake of LDLC in cells was detected by DiI-LDL. Real-time fluorescence quantitative PCR and Western blot were used to detect sterol regulatory element binding protein-2 (SREBP-2) and low density lipoprotein receptor (LDLR) expressions at mRNA and protein level, respectively. LDLR content in the hepatic cell membrane was detected by flow cytometry. Results Ezetimibe reduced the content of TC and LDLC in the plasma of mice, increased the content of HDLC and reduced the lipid deposition in liver. Ezetimibe enhanced the ability of HepG2 cells to take LDLC, and promoted the transformation of FC into CE. Ezetimibe up-regulated the expressions of SREBP-2 and LDLR in HepG2 cells, and increased the distribution of LDLR on the cell membrane. Conclusion Ezetimibe can promote the uptake of LDLC in liver cells by up-regulation of SREBP-2 and LDLR.