Aim To observe the change of serum total cholesterol （TC） and efficiency of the synthesis and absorption of cholesterol in the mfat-1 mice after high fat diet, and to explore the relative mechanism. Methods 12 mfat-1 mice as experimental group and 12 C57BL / 6 mice as control group were given high fat diet for 4 weeks. Blood was taken from the heart, and the liver tissue extracted four weeks later. The content of liver lipids eicosapentaenoic acid（EPA）, docosahexenoic acid（DHA）, docosapentaenoic acid（DPA）,α-linolenic acid（ALA）, linoleic acid（LA）, arachidonic acid（AA）, serum TC, and the level of cholesterol synthesis markers （squalene, desmosterol） , absorption markers（campesterol,β-sitosterol,stigmasterol） , were determined with gas chromatogram. The express of cholesterol synthesis rate-limiting enzyme in liver, 3-Hydroxy-3-methylglutaryl- coenzyme A reductase（HMG-COAR）, were determined with ELISA. Results In the contents of liver membrance lipid, the ratio of ω-6/ω-3 polyunsaturated fatty acids in the experimental group was significantly lower than that in the control group （1.70 vs 6.04, P<0.01）. The levels of liver HMG-COAR in the experimental group were significantly lower than those in the control group（379.76±22.37 vs 443.98±27.99, P<0.01）. The levels of plasma TC, cholesterol synthesis markers （squalene, desmosterol） and absorption markers （campesterol,β-sitosterol, stigmasterol） in the experimental group were significantly lower than those in the control group （0.91±0.07 vs 1.16±0.85, P<0.01 0.63±0.17 vs 1.24±0.36, 3.89±0.54 vs 6.57±1.06, P<0.05 16.10±2.28 vs 21.24±2.96, 1.07±0.18 vs 1.76±0.43, 28.78±5.60 vs 42.30±5.81, P<0.01, respectively）.Conclusion The mfat-1 mice can inhibit cholesterol synthesis and absorption, thus decrease the total cholesterol.