Effect Of Transient Expression Of Human Cholesteryl Ester Hydrolase On The Cholesterol Metabolism In RAW 264.7 Macrophage

ObjectiveTo study the effect of transient expression of human cholesteryl ester hydrolase(hCEH) on the cholesterol metabolism in RAW 264.7 macrophages.MethodsRAW 264.7 macrophages were transfected transiently with pEGFP-N1-hCEH under the stimulation of FuGENE HD. After 24 h, macrophages were loaded with oxidized low density lipoprotein(100mg/L) for 48 h.The expression of pEGFP-N1-hCEH was detected with GFP fluorescence and Western Blot. The cellular lipid accumulation was examined by oil red O staining. High performance liquid chromatography was used for qualitative and quantitative analysis of intracellular cholesterol and cholesterol esters.ResultsThere were the most number of fluorescent cells after 72 h. The transfection rate was about 40%. The protein expression level of human CEH significantly increased and partly inhibited expression of mouse CEH protein in mouse macrophage.Compared with ox-LDL group, oil red O-positive cells of hCEH transfection group were greatly reduced.The cellular contents of total cholesterol, free cholesterol and cholesterol ester of hCEH transfection group were also obviously decreased. ConclusionsThe gene of hCEH expresses in mouse macrophages cell line RAW 264.7, and inhibits atherosclerotic foam cell formation through reducing intracellular cholesterol level.