This study aimed to determine the fermented milk of Lactobacillus casei Shirota strain potential on cholesterol levels, SOD activity, LPL enzyme activity, and cardiac histopathology. This study used 25 rats (Rattus novergicus) Wistar strain aged 2-3 months, weighing 150-200 grams. These rats were divided into 5 groups: negative control group, positive control group with high-cholesterol diet for 21 days, and therapy group with 1.0 ; 1.5 ; 2.0 mL/rat/day of fermented milk for 14 days. The analysis methods used in this study are One Way ANOVA and Tukey. The results showed that the fermented milk of Lactobacillus casei Shirota strain had a strong effect on blood cholesterol levels and increasing the SOD activity. This study also showed that the fermented milk of Lactobacillus casei with 2.0 mL/rat/day is the most effective dose to decrease the total cholesterol and increase the SOD activity.Keywords: cholesterol, fermented milk of Lactobacillus casei Shirota strain, total cholesterol, SOD INTRODUCTIONCholesterol is a steroid alcohol on the class of lipids (fats) which play a part as a source of calories. In addition, cholesterol is a component of fat found in the blood of animals and humans that produced by the liver and used as an energy producer. It is also the wallforming cells in the body and as a basic ingredient of the steroid hormones formation in the body. Cholesterol is the precursor of all steroids, such as corticosteroids, sex hormones, bile acids, and vitamin D.Consuming lots of high-cholesterol diet can lead to the increasing of cholesterol levels. It causes hypercholesterolemia, atherosclerosis in the arteries, and blood vessels constriction in the heart, brain, kidneys, and eyes when it reaches beyond the normal limits. Hypercholesterolemia is a condition when the blood cholesterol level exceeds the normal value of 200 mg/dL. In the body, cholesterol found in lipoproteins, such as chylomicrons, which contains triglycerides, VLDL (very low density lipoprotein) to transport triglycerides. The triglycerides formed by the heart, LDL (low density lipoprotein) or beta-lipoprotein consisting of 25% protein and 40% cholesterol and HDL (high density lipoprotein) or alpha lipoprotein contain 50% protein and 20% cholesterol [1].