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Updated: Jun 7, 2026

Oxygen-Glucose Deprivation and Reoxygenation as an In Vitro Ischemia-Reperfusion Injury Model for Studying Blood-Brain Barrier Dysfunction
Published on: May 7, 2015
[Reactive oxygen species participates in low glucose-induced injury of ECV304 cells]
Ning Liu1, Yao-ming Xue, Bo Zhu
1Department of Endocrinology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. ning_2392@ hotmail.com
Objective:
To investigate the mechanism of low glucose-induced injury in human ECV304 cells.
Methods:
Human umbilical vein endothelial cell line ECV304 were stimulated with low concentrations of glucose. The level of reactive oxygen species (ROS) in cells was detected at different time points within 12 h by kinetic measurement of dichlorofluorescein (DCF) fluorescence produced by oxidation of an oxidant-sensitive dye 2,7-dichlorofluorescein (DCFH). ECV304 cell viability was assessed with MTT assay and NADPH oxidase activity detected using lucigenin-enhanced chemiluminescence assay following cell stimulation with low glucose and apocynin.
Results:
Low-glucose exposure of ECV304 cells time- and dose-dependently induced ROS production, and which was decreased by apocynin treatment. Apocynin pretreatment of the cells inhibited ROS production by 44% in cells exposed to 2.8 mmol/L glucose and by 60% in cells without glucose exposure.
Conclusions:
Low glucose of ECV304 cells induces ROS production to cause cell injury, which is mediated partially by NADPH oxidase activation.
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