Endothelial cells have many important roles in the cardiovascular system, such as controlling vasomotor actions and hemostasis. In the event of endothelial cell dysfunction, the risk of cardiovascular disease increases. Therefore, the objective of this study was to investigate the early detection and diagnosis of endothelial cell dysfunction. Injury and restoration in vascular endothelial cells exposed to ischemic stress may affect changes in the electrical impedance. We measured the status of the endothelial cell layer by using microelectrochemical impedance spectroscopy. We used cultured rat primary vascular endothelial cells to measure the electrical impedance under different conditions (control, ischemia, and recovery). Our results revealed that the electrical impedance in vascular endothelial cells under different conditions has quantitatively distinct values. At the optimal frequency, the real parts (Z) of the impedances for the control group, ischemic group, and recovery group were 0.54 kΩ 0.28 kΩ and 0.58 kΩ respectively. The imaginary parts (Z) of the impedances for each group were − 0.19 kΩ − 0.12 kΩ and − 0.18 kΩ respectively. The values for both the recovery group and control group were similar. In this context, electrical impedance measurement could be considered as possible method for direct detection of vascular endothelial cell injury in ischemic conditions. To the best of our knowledge, this study is the first attempt to measure changes in the electrical impedance of vascular endothelial cells during ischemic damage and the recovery processes.