Background: Gestational diabetes mellitus (GDM) is any degree of glucose intolerance diagnosed during pregnancy. Metabolic abnormalities in GDM are associated with pancreatic β-cell destruction and decreased insulin sensitivity. Carbohydrate-responsive element-binding protein (ChREBP) and sterol regulatory element-binding protein-1c (SREBP-1c) are two transcription factors that play a key role in carbohydrate and lipid metabolism homeostasis. Our aim in this study was to elucidate the role of these proteins in GDM. Methods: The study included 50 pregnant women diagnosed with GDM and 52 pregnant women with normal glucose tolerance. Maternal blood samples were collected from pregnant women between the 24th and 28th weeks of pregnancy. Serum SREBP-1c and ChREBP levels were measured with commercial ELISA kits according to the manufacturer’s instructions. Results: Serum ChREBP (p=0.044) and SREBP1c (p=0.042) levels of pregnant women with normal glucose tolerance were found to be statistically significantly higher than women with GDM.