Purpose: In this study, we intend to explore the possible mechanism of statins on endothelial cell dysfunction caused by high glucose. Methods: Internet database was used to identify the potential targets and signal pathways of statins against endothelial dysfunction in diabetes. We observed the changes in cell survival, proliferation, apoptosis, and NO level in human umbilical vein endothelial cells (HUVECs) with high glucose and rosuvastatin. In addition, the genes and proteins expression of eNOS, CLDN-1, OCLN, ZO-1, HIF-1, VEGF, and Bax were detected in HUVECs. Results: The results of network pharmacology and molecular docking indicate that HIF1A may be a key target of rosuvastatin in improving endothelial injury under high glucose environment. High glucose caused the cell apoptosis, decreased the NO production, reduced the expression of tight junction-related proteins, and inhibited the expression and effect of HIF-1α on HUVECs. However, rosuvastatin was able to reverse these effects. Effects of rosuvastatin on vascular endothelial cell dysfunction induced by high glucose was abolished by the HIF-1 inhibitor YC-1. Conclusion: Our study demonstrates that rosuvastatin might act on HIF-1 molecules, enhance cell survival, and decrease apoptosis under the high glucose condition in HUVECs.