Hashimoto's thyroiditis is an autoimmune disorder that primarily affects the thyroid gland, leading to chronic inflammation and impaired thyroid function. On the other hand, transforming growth factor beta-1 (TGF-β1) is a protein that plays a crucial role in regulating various cellular processes, including immune responses and tissue remodelling. The objective of this study was to investigate the association of TGFβ-1 levels and its gene polymorphisms in Hashimoto's Thyroiditis (HT) patients and the immune system attack of the thyroid gland cells, by the presence of antibodies, including thyroglobulin antibody (TG-AB) and thyroid peroxidase antibody (TPO-AB), which serve as biochemical markers for Hashimoto's Thyroiditis. A case-control study encompassed a cohort of 100 female subjects within the age range of 20 to 53 years, the study consisted of a group of diagnosed Hashimoto's thyroiditis patients and an equivalent number of healthy volunteers who were carefully selected to match the age and gender characteristics of the patient group. The study employed allele-specific arm-PCR technique to examine the genotype of TGF-β1 gene polymorphisms. Furthermore, levels of T3, T4, TSH, fT3, fT4, Anti TPO, and Anti-TG were quantified using the ELISA method, utilizing specific commercial kits designed for analysis within the respective study groups. Current work showed a highly significant variation in TGFβ1 levels in HT patients group when compared with healthy group, in addition to these findings suggest that the presence of the CG, GG genotype of rs1800471 and the CT, TT genotypes in rs1982073 are associated with an increased susceptibility to HT disease. The present study concluded that genotypes distribution and associated risks highlight the potential role of this genetic variation in the development and manifestation of HT disease.