The dynamic changes of reactive oxygen species expression in rat brain tissue following focal cerebral ischemia/reperfusion

doi:10.3969/j.issn.1006-7795.2015.05.006

Abstract

Abstract: Objective To investigate the dynamic changes of reactive oxygen species (ROS) during reperfusion time in a rat middle cerebral artery occlusion (MCAO)/reperfusion model and to investigate the role of ROS in cerebral ischemic injury. Methods Twenty-five male Sprague Dawley rats were divided randomly into 5 groups: sham group and MCAO group (90 min ischemia with 3,6,12 or 24 h reperfusion). MCAO operation was performed by using suture method. The rats underwent right MCAO for 90 min. The mean arterial blood pressure (MABP) and rectal temperature were monitored to keep in normal range during the operation. Brains were removed after the animals were euthanized at 3, 6, 12 and 24 h after reperfusion respectively and dyed with 2,3,5-triphenyltetrazolium chloride (TTC) to measure the infarct volume. Cryostat sections were prepared on a freezing microtome. ROS-specific staining and H₂DCF-DA fluorescent staining were used to detect the levels of ROS in the penumbra of the brain. Results 1) There was no statistically significant difference in MABP and rectal temperature between sham and MCAO groups. 2) Sham group rats had no penumbra in the brain where little H₂DCF-DA-positive cells were observed. Lots of H₂DCF-DA-positive staining cells were observed in the penumbra of MCAO group rats at 3, 6, 12 and 24 h after reperfusion. From 3 h after reperfusion, the numbers of H₂DCF-DA-positive staining cells in the penumbra of MCAO group rats showed reperfusion-time-dependent increasing tendency, with significant difference between 3 h and 6 h, as well as 12 h and 24 h (P<0.05). 3) The result of TTC staining showed that no infarction was observed in the brain of sham group rats. The infarct volume of MCAO group rats showed reperfusion-time-dependent increasing tendency (from 3 h to 24 h after reperfusion, P< 0.05). 4) The number of H₂DCF-DA-positively stained cells numbers showed positive correlation with the volume of brain infarction in MCAO group (r=0.833,P<0.01). Conclusion ROS level was increased along with the extension of reperfusion time in a rat focal cerebral ischemia/reperfusion model. ROS might play an important role in cerebral ischemic/reperfusion injury.

Key words: reactive oxide species (ROS), cerebral ischemia, middle cerebral artery occlusion, reperfusion, rat

CLC Number:

R743.31

Yan Feng, Zhao Yongmei, Luo Yumin, Yin Jie, Li Sen, Fang Yalan, Liu Kejian. The dynamic changes of reactive oxygen species expression in rat brain tissue following focal cerebral ischemia/reperfusion[J]. Journal of Capital Medical University, 2015, 36(5): 694-698.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2015.05.006

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2015/V36/I5/694

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