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Background: In the present study, the protective effect of vitamin D3 against sodium fluoride induced nephrotoxicity was evaluated in rats. A vital role in kidney protection and ensuring the longevity of healthy kidneys is played by vitamin D3. The risk of autoimmune disorders, internal diseases, cancer, and infections is increased when vitamin D3 deficiency is present. Objective: a consequence of prolonged exposure to numerous pharmaceutical products, including industrial material, fluoride-containing herbicides, medications, and fire extinguishers. Concerns concerning fluoride toxicity have grown recently. As a result, scientists are constantly hunting for medications that could reduce toxicity in different human organs. This study will evaluate the nephroprotective effects of vitamin D3 against renal damage caused by sodium fluoride. Methods: In this study, 30 rats were divided into 5 groups of 6 rats each. Rats were fed water containing 100 ppm of sodium fluoride, and vitamin D3 was administered orally for 30 days in three different doses: 250 IU per kg, 500 IU per kg, and 1000 IU per kg. The Nephrotoxicity indicators were assessed. Results: Sodium fluoride considerably decreased total protein while dramatically increasing urea, uric acid, and creatinine. It also helps people lose weight. On the other hand, in the test group, vitamin D3 significantly decreased levels of urea, uric acid, and creatinine while also raising levels of total protein. Furthermore, it enhanced the rats' body weight in test groups. Results: The test animals that drank 100 parts per million water for 30 days had high levels of urea, uric acid, and creatinine in their blood, but lower levels of total protein. The test group received vitamin D3 doses of 250, 500, and 1000 IU/kg, leading to weight gain, decreased urea, uric acid, and creatinine levels, and increased levels of total protein. The study group that received 1000 IU/kg performed well. Vitamin D3 considerably reduced the nephrotoxicity brought on by sodium fluoride in a dose-dependent manner.