Although negative evidences of trans fatty acids(tFAs) are gradually increasing, the hypercholesterolemic effect of tFAs are controversial and its biological significances are still not known. The objective of this study was to examine the effect of dietary tFAs on cholesterol metabolism and membrane fluidity in rats. Animals were fed diets containing 0.5% cholesterol and 20% test fats(margarine, butter, corn oil) for 8 weeks. Each test fats(margarine, butter, corn oil) contained 25%, 8.5% and no tFAs, respectively, Plasma total triglyceride(TG) were not different among diet treatments. Feeding trans fat diets (margarine and butter) lowered plasma cholesterol. Specially, butter diet elevated LDL-cholesterol and decreased HDL-cholesterol levels, resulting in the highest atherogenic index among diet treatments. Hepatic cholesterol concentration and HMG CoA reductase activity were also decreased, whereas fecal excretion of cholesterol was increased in trans fat-fed animals. trans fat containing diets also decreased hepatic membrane fluidity. From these results, it can be concluded that hypercholesterolemic effect of tFAs can be modulated by the fatty acid composition of diets, but its spatial cofiguration may help behave like saturated fatty acid in membrane fluidity.