Background: There is inconsistent data on the effects of vitamin D supplementation on metabolic profiles in pregnant women with gestational diabetes mellitus (GDM). The aim of the current study was to assess the effect of vitamin D supplementation on the glycemic and metabolic status of pregnant women with GDM.Methods: In this randomized double-blind controlled trial pregnant women with GDM referred to the Diabetes Clinic of Hormozgan University of Medical science, Bandar Abbas, 2016-2017, were randomly assigned to the intervention (n=25) and placebo (n=25) groups. Subjects in the intervention group received two 50000 IU vitamin D3 pearls (at baseline and at day 21) plus 1000 mg calcium daily, and those in the placebo group received 2 placebos at the same times and 1000 mg calcium daily. Fasting blood samples were collected at baseline and after 6 weeks for the measurement of lipid profile, fasting plasma glucose (FPG), insulin, 25-hydroxy vitamin D, calcium, and phosphorus. Homeostatic model assessment for insulin resistance (HOMA-IR) was calculated.Results: Serum 25-hydroxy vitamin D significantly increased in the intervention group (P<0.001). Vitamin D supplementation significantly decreased FPG (P=0.049), serum insulin level (P=0.001), and HOMA-IR (P=0.001) in the intervention group compared to the placeb group. Furthermore, vitamin D significantly reduced serum triglyceride (P=0.0.002), whereas it had no significant effect on total cholesterol (P=0.099), low-density lipoprotein cholesterol (LDL-C) (P=0.279) , and high-density lipoprotein cholesterol (HDL-C) (P=0.472) compared to the placebo group.Conclusions: Vitamin D supplementation can be beneficial in the regulation of FPG, insulin level, and HOMA-IR in pregnant women with GDM.Trial registration: The research has retrospectively been registered at the Iranian Registry of Clinical Trials (IRCT) with the registration number: IRCT20150607022585N3 and is accessible at the following website: www.irct.ir.