The purpose of the present study was to examine the relationship between the changes in the expressions of several mRNAs and changes in endothelial function in streptozotocin-induced diabetic and chronic short-term insulin-treated rats. Aortas from later-stage (10 week) diabetics, but not those from their insulin-treated counterparts, showed an impaired endothelial function. We found that the mRNA expressions for 30 genes were significantly upregulated, while those for 13 other genes were downregulated in aortic endothelial cells from diabetes. In later-stage diabetes, chronic insulin treatment ameliorated the endothelial dysfunction and normalized the expressions for 20 out of the 43 genes altered in diabetes. Further, 12 of the remaining 23 genes were altered by high-dose insulin treatment in the controls. In early-stage (1 week) diabetic aortas, which did not show impaired endothelial function, expression changes were shown by only 12/30 and 5/13 of the genes up- or downregulated, respectively, in later-stage diabetes. Thus, in the diabetic aortas endothelial gene expressions and function exhibited time-related changes, and several gene expressions and endothelial function were normalized by insulin treatment. The hyperinsulinemia caused by this treatment may oppose the alterations in some gene expressions and the endothelial proliferation (cell growth-related gene expressions) that occur in established diabetes.