Abstract: The role of probiotics and prebiotics as cholesterol reductive agents has drawn a great deal of attention. However, there have been few studies, investigating the effect of prebiotics and probiotics together on lipid profile. The purpose of study was to evaluate the effect of probiotic cheese (PCh) and chicory root extract consumption on lipid profile in a large sample. 180 subjects aged 18–65, with <6 mmol/l total cholesterol participated in the 7-week trial. Subjects randomly assigned into three 60-person groups. E1 consumed 30 g of PCh plus chicory root extract (daily), E2 consumed 30 g of PCh (daily) along with a control group. Probiotics strains in products were Lactobacillus acidophilus LA5 and Bifidobacterium lactis BB12. Anthropometric measurements and blood sampling were conducted at the baseline and end of trial. There was a significant reduction within both intervention groups throughout the study: intervention 1, (LDL: −0.79, 95 % CI −0.96, −0.61, P < 0.0001; cholesterol: −1.02, 95 % CI −1.41, −0.63, P < 0.0001; triglyceride: −0.59, 95 % CI −0.67, −0.51, P < 0.0001; HDL: 0.44, 95 % CI, 0.37, 0.51, P < 0.0001); intervention 2 also showed significant improvements in the mentioned factors. Comparing results of both groups with control showed significant improvements in all factors except triglyceride of intervention 2. PCh and chicory extract showed great effects on improvements of lipid profile, thus they can be used for prevention and treatment purposes.