Aim To investigate the effects of calculus bovis cultivated by glucuronidase （CBCG） on high density lipoprotein （HDL） anti-atherosclerotic functions in mice fed with high fat high cholesterol diet. Methods Forty one male 7-8-week-old C57BL/6J mice fed with high fat diet （15.8% fat and 1.25% cholesterol） were randomly divided into three groups: control group （n＝14）, medium-dose （0.75 g/（kg·d）, n＝13） and high-dose （2.25 g/（kg·d）, n＝14） CBCG treated groups. After 12 weeks of treatment, blood was collected from the retroorbital sinus of the C57BL/6J mice without dietary exposure for 12 h. LDL and HDL₃ were separated by sequential ultracentrifugation. HDL functionality assay, Cu²⁺-induced LDL oxidation assay, endothelial cell-monocyte adhesion assay and MTT assay were determined to analyse the functional properties of the HDL particle. Forty male 7-8-week-old apolipoprotein E knockout mice fed with high fat diet （15.8% fat and 1.25% cholesterol） were randomly divided into four groups: control group, low-dose （0.25 g/（kg·d））, medium-dose （0.75 g/（kg·d））, and high-dose CBCG （2.25 g/（kg·d）） treated groups. After 8 weeks of treatment, blood was collected from the retro-orbital sinus of apolipoprotein E knockout mice without dietary exposure for 12 h. Plasma concentrations of IL-6 and TNF-α were determined by ELISA. Plasma levels of malondialdehyde （MDA） and the activity of paraoxonase-1 （PON-1） were determined by spectrophotometric method. The bioactive compounds, namely bilirubin and taurine were determined by ELISA. Plasma concentrations of total cholesterol （TC） and HDLC were determined by enzymatic methods. Non-HDLC was calculated as TC minus HDLC. Results The anti-atherosclerotic actions of CBCG were linked with improving the functional quality of HDL particle in C57BL/6J mice, including reverse cholesterol transport （RCT）-promoting, anti-oxidative （prevention of LDL oxidation）, endothelial protective as well as anti-inflammatory （induce adhesion of monocyte to human umbilical vein endothelial cells） properties. At the end of eight weeks of intragastric administration of CBCG, ELISA revealed that the production of tumor necrosis factor-α and interleukin-6 were significantly suppressed. Spectrophoto-metric measurement showed that plasma levels of MDA were significantly decreased and the activity of PON-1 was significantly improved. CBCG significantly increased the plasma level of bilirubin while the change of taurine level was not obvious. Plasma analysis by enzymatic method showed that CBCG remarkably increased plasma HDLC in mice fed with high fat high cholesterol diet. Conclusions The results reveal that CBCG significantly improve the functional quality of HDL particle in C57BL/6J mice, including RCT-promoting, anti-oxidative, endothelial protective as well as anti-inflammatory properties. The improving function may be linked with the anti-oxidation of CBCG in plasma.