Aim To investigate the effect of Ezetimibe on cholesterol accumulation in loaded-lipid cells derived from vascular smooth muscle cells(VSMC).Methods Loaded-lipid cells model were established by 20 mg/L Chol∶MβCD treated VSMC.Loaded-lipid cells were respectively treated with different concentration of ezetimibe(0,3,10,30 μmol/L) for 24 h,and with 30 μmol/L of ezetimibe for different time(0,6,12,24,48 h).The concentration of total cholesterol(TG) and free cholesterol(FC) were detected by high performance liquid chromatography(HPLC).Cholesterol accumulation in cells were observed by oil red O staining.Results(1) Intra-cellular TG were110.8±3.56,90.5±.2.30,60.9±1.86 mg/g and FC were 85.5±3.12,74.6 ±2.89,53.8±1.56 mg/g,respectively with loaded-lipid cells treated with different concentration of ezetimibe.Compared with model group,there was significant difference in medium-dose and high-dose groups(P<0.05),TG and FC had a reducing trend with rising drug concentration(tendency χ2=11.56,P=0.007) and the best drug concentration was 30 μmol/L.(2)Treated loaded-lipid cells for different time(0,6,12,24,48 h) with 30 μmol/L ezetimibe,the content of TG were 120.1±2.37,117.9±3.05,80.3±.2.81,69.2±1.56,72.1±2.98 mg/g and FC were 92.9±2.11,90.7±3.76,71.4±2.57,60.1±1.21,64.2±2.48 mg/g respectively.TG and FC gradually decreased with treated time,and both of them had negative correlations with treated time(r=-0.776,P<0.05 and r=-0.786,P<0.05 respectively).Compared with 0 h group,there was significant difference for FC in 12 h,24 h and 48 h groups(P<0.01).There was time-depended tendency for ezetimibe and it was best for 24 hours.Conclusion Ezetimibe decrease cholesterol accumulation in vascular smooth muscle cells.