Сholelithiasis is a multifactorial process that is influenced by both environmental and genetic factors. Some evidence suggests that total plasma homocysteine correlates with the presence of gallstones, suggesting that hyperhomocysteinemia is a risk factor for cholelithiasis. The aim of this work was to analyze the association of polymorphic variants of the methylenetetrahydrofolate reductase MTHFR (rs1801133 (677C > T), rs1801131 (1298A > C)) and methionine synthase reductase MTRR (rs1801394 (66A > G)) genes with the development of gallstone disease in individuals from the Republic of Bashkortostan. DNA samples from 196 patients with cholelithiasis and 274 healthy individuals aged 23-87 years living in the Republic of Bashkortostan were used as research material. Genotyping was performed using the real-time PCR method. It has been established that the rs1801133*T allele and the rs1801133*TT genotype of the MTHFR gene are markers of an increased risk of developing cholelithiasis. An association was established between the rs1801133*TT genotype of the rs1801133 polymorphic variant of the MTHFR gene and the moderate severity of cholelithiasis and hereditary burden in patients with cholelithiasis. A study of the polymorphic variant of the MTRR gene revealed that the rs1801394*G allele increases the risk of cholelithiasis. Analysis of associations of the polymorphism rs1801131 of the MTHFR gene with the development of cholelithiasis did not reveal statistically significant differences between the compared groups of patients and controls. Determination of homocysteine levels and genetic testing of MTHFR and MTRR polymorphisms in patients with cholelithiasis may be useful in clinical practice.