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Updated: Jun 5, 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
[Study of DNA damage with neuronal ischemia/reperfusion in vitro]
Yu-Min Luo1, Zhen Qin, Jun Chen
1Department of Neurology, Huashan Hospital of Shanghai Medical University, Shanghai 200040, China.
Aim And Methods:
To investigate DNA strand damage in a model of cerebral ischemia/reperfusion injury in vitro, model of ischemia/reperfusion was produced by incubating the primary neuronal cultures to various durations of hypoxia and glucose deprivation (HGD). DNA single and double strand breaks were detected using PANT and TUNEL staining respectively.
Results:
A few cell death occurred after 2 h of HGD. Less than 30 percent of cell died after 4 h of HGD, whereas 6-8 h of HGD resulted in cell death in 80 percent of neurons. Neuronal cell death reached the peak 10-18 h after 6 h of HGD, while it took as early as 2 h after 8 h of HGD. Following HGD, PANT positive cells were remarkably increased, which was proportional to the duration of HGD. At 5 minutes after 2, 4, 6 or 8 h of HGD, PANT positive cells were 30%, 50%, 80%, 90% respectively. Meanwhile, no determined in another cultures, DNA double strain breaks were not significantly increased.
Conclusion:
DNA strain breaks is a very early event of DNA damage following in vitro ischemia/reperfusion, especially DNA single strain breaks.
