A rat model of diabetic artery calcification.

Objectives: The aim of this study was to develop a rat model that shares the similarities of calcification in diabetes. Methods: Male Wistar rats received a high-fat diet during 8 weeks followed by a low dose of streptozotocin. Half of them were treated with vitamin D3 and nicotine (VDN) 1 week after streptozotocin injection (DM) (DM+VDN). The others were treated with vehicle (DM). Arterial calcification was facilitated by vitamin D3 and nicotine in age-matched rats (VDN). Measurements of metabolic parameters, aortic calcium content, von Kossa staining, alkaline phosphatase activity and immunohistochemistry for osteopontin were performed. The expression of genes and proteins associated with calcification were also examined. Results: Treatment with VDN alone resulted in a small but not significant elevation of calcium content in aorta. However, in DM+VDN, aortic calcium content increased significantly as compared to the VDN. This calcification had also dramatically increased, as shown by von Kossa staining. Mechanistic studies revealed that addition of VDN treatment in diabetic rats enhanced both activity and mRNA expression of alkaline phosphatase. This treatment in diabetic-rats also enhanced the expression of core binding factor α 1 and its downstream protein osteopontin. In aorta, strong immunostaining for osteopontin was observed in DM+VDN. The result was also confirmed by Western blot analysis. Conclusion: These results suggest that a model of accelerated arterial calcification in diabetes have been established and this model could be useful to investigate mechanisms related to vascular complication in diabetes.