Toll-like receptor 4 (TLR4) gene plays important roles in the susceptibility to type 2 diabetes mellitus (T2DM). This study aims to detect the potential association of TLR4 gene polymorphisms with the susceptibility to T2DM in the Chinese Han population. 685 T2DM patients and 690 healthy controls were enrolled in this case–control study. The genotypes of TLR4 gene polymorphisms were analyzed by the polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing methods. We detected the g.13726T > C and g.15090G > A genetic polymorphisms. Our data supported that the g.13726T > C and g.15090G > A genetic polymorphisms were significantly associated with the increased susceptibility to T2DM in the homozygote comparison, recessive model, and allele contrast (all P-values < 0.01). The allele-C and genotype-CC of g.13726T > C and allele-A and genotype-AA of g.15090G > A genetic polymorphisms might be the risk factors for increasing the susceptibility to T2DM. These preliminary findings indicate that the g.13726T > C and g.15090G > A genetic polymorphisms of TLR4 gene are potentially related to the susceptibility to T2DM in the studied population, and might be used as molecular markers for evaluating the risk of T2DM.