Edaravone protects the neural stem cell proliferation under hypoxia in vitro

LEI Shan; ZHANG Pengbo; LI Weisong; GAO Ming; HE Xijing; ZHENG Juan; LI Xu; WANG Xiao; WANG Ning; ZHANG Junfeng; QI Cunfang; LV Haixia; CHEN Xinlin; LIU Yong

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Edaravone protects the neural stem cell proliferation under hypoxia in vitro

LEI Shan 1,ZHANG Pengbo 2 *,LI Weisong 2,GAO Ming 2,HE Xijing 3,ZHENG Juan 2,LI Xu 2,WANG Xiao 2,WANG Ning 2,ZHANG Junfeng 4,QI Cunfang 4,LV Haixia 4,CHEN Xinlin 4,LIU Yong 4

1. Department of Anesthesiology, Second Affiliated Hospital of Xi’an Jiaotong University School of Medicine, 157# West 5 road, Xi’an 710004

2.Department of Anesthesiology, Second Affiliated Hospital of Xi’an Jiaotong University School of Medicine, 157# West 5 road, Xi’an 710004

3.Department of Orthopedics, Second Affiliated Hospital of Xi’an Jiaotong University School of Medicine, 157# West 5 road, Xi’an 710004

4.Institute of Neurobiology, National Key Academic Subject of Physiology of Xi’an Jiaotong University School of Medicine, 76# Yanta West Road, Xi’an 710061

*Correspondence author

#Submitted by

Subject:

Funding: The Specialized Research Fund for the Doctoral Program in Higher School of China （No.20100201110051）, the National Natural Science Foundation of China （No.No. 8107107）, the Program for New Century Excellent Talents in University of China （No.NCET-08-0436）

Opened online:16 January 2014

Accepted by: none

Citation: LEI Shan,ZHANG Pengbo,LI Weisong.Edaravone protects the neural stem cell proliferation under hypoxia in vitro[OL]. [16 January 2014] http://en.paper.edu.cn/en_releasepaper/content/4581610

Hypoxia stimulates neural stem cell (NSC) proliferation and enhances generation of endogenous reactive oxygen species (ROS). However, the role of edaravone in NSC proliferation under severe hypoxia remains unclear. We used a free radical scavenger edaravone to investigate the effect and underlying mechanism of edaravone on NSC proliferation under 0.3% oxygen condition. Neural stem cells (NSCs) were prepared from 14-day embryonic mice and identified by immunohistochemical analysis. NSCs were incubated with edaravone (0μM or 100 μM) for 24 h under hypoxia (0.3% oxygen) or normoxia (5% CO2/95% air), respectively. Cell proliferation was evaluated by 5'-bromo-2'-deoxyuridine (BrdU) incorporation. ROS were detected by 2,7-dichlorofluorescien diacetate (DCF-DA) assay. Cell apoptosis was assessed by flow cytometric analysis and terminal dUTP nick-end labeling. HIF-1α and activated caspase-3 proteins were quantified by western blot analysis. After 24 hours of hypoxia, most cells were nestin and HIF-1α positive. Hypoxia increased ROS generation, elevated protein levels of HIF-1α and activated caspase-3, and facilitated NSC apoptosis compared with normoxia (P < 0.01), respectively. Treatment with edaravone, a free radical scavenger, inhibited significantly the above changes induced by hypoxia (P < 0.01). Edaravone also increased significantly NSC proliferation under hypoxia (P < 0.01). These findings indicate that edaravone promotes NSC proliferation under 0.3% oxygen condition probably by inhibiting HIF-1α, caspase -3 and apoptosis under severe hypoxia. The present data reveal a novel ROS-dependent HIF-1α-mediated caspase-3 activation pathway which leads to NSC apoptosis in vitro.

Keywords:neurobiology; hypoxia; neural stem cells; proliferation; reactive oxygen species; hypoxia inducible factor -1 alpha

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