Cardiovascular diseases caused by atherosclerosis due to hypercholesterolemia have emerged as a significant threat to human health. Recently, probiotics exhibiting cholesterol-lowering properties have emerged as a prominent area of research. Numerous studies have demonstrated that Lactobacillus reuteri can effectively reduce endogenous cholesterol synthesis, regulate cholesterol transport, and promote cholesterol degradation by modulating the expression of key genes such as sterol-regulatory element binding protein 2 (SREBP-2), 3-hydroxy-3-methyl-glutaryl coenzyme A reductase (HMGCR), and cholesterol 7 alpha-hydroxylase (CYP7A1) in both the liver and intestinal epithelial cells of the host. Consequently, this leads to a notable decrease in total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C）levels in the host serum. The present paper offers a comprehensive overview of the underlying mechanisms responsible for the cholesterol-lowering effects exerted by Lactobacillus reuteri, aiming to provide valuable insights into the treatment of hypercholesterolemia and the development of probiotics with cholesterol-lowering properties.