Diabetes was shown to be associated with a considerable lowering of 25(OH)D3 in blood serum of mice. Vitamin D3 deficiency was correlated with impaired mineral metabolism in bone tissue, indicating the development of secondary osteoporosis. A decrease in weight, length and diameter (diaphysis, proximal metaepiphysis) of tibia in diabetic animals was observed as compared with control. Diabetes caused hypocalcemia, hypophosphatemia and increased enzymatic activity of alkaline phosphatase (ALP) and its isoenzymes in serum. This changes were accompanied by the impairments of vitamin D3 25-hydroxylase isoforms (CYP27A1 and CYP2R1) expression, which are the main enzymes of cholecalciferol biotransformation to 25(OH)D3 – precursor of hormonally active form of vitamin D3. A decrease in bone resorption processes was established after vitamin D3 administration as it is evident from normalization of bone morphometrical parameters and mineral metabolism in diabetic mice. Vitamin D3 ability to counter diabetes-induced alterations in bone tissue can be ascribed, at least in part, to its positive effects on the formation of vitamin D3 hormonally active forms.