Astrocyte-conditioned medium attenuates glutamate-induced apoptotic cell death in primary cultured spinal cord neurons of rats

被引:12
作者
Lu, Xiaojian [1 ]
Al-Aref, Rami [2 ]
Zhao, Dong
Shen, Jianhong [1 ]
Yan, Yaohua
Gao, Yilu [1 ]
机构
[1] Nantong Univ, Affiliated Hosp, Dept Neurosurg, Nantong 226001, Jiangsu, Peoples R China
[2] Wayne State Univ, Sch Med, Dept Neurol, Detroit, MI 48201 USA
基金
中国国家自然科学基金;
关键词
Astrocyte-conditioned medium; Glutamate; Apoptosis; Spinal cord neurons; IMPACT INJURY; BRAIN; ACTIVATION; CYTOKINES; ACID;
D O I
10.1179/1743132815Y.0000000059
中图分类号
R74 [神经病学与精神病学];
学科分类号
摘要
Objectives: To better understand the neuroprotective role of astrocytes in spinal cord injury (SCI), we investigated whether astrocyte-conditioned medium (ACM) can attenuate glutamate-induced apoptotic cell death in primary cultured spinal cord neurons. Methods: Spinal cord neurons were pretreated with ACM for 24 hours. Subsequently, they were exposed to glutamate (125 mu M) for 1 hour. The neurons were then incubated for 24 hours. Following that, measurements assessing cell viability and lactate dehydrogenase (LDH) release were performed. Apoptosis was confirmed through cell morphology using Hoechst 33342 staining and terminal deoxynucleotidyl transferase dUTP-mediated nicked end labeling (TUNEL) assay. Assessment for expression of apoptotic enzymes, including Caspase-3, Bcl-2 and Bax, was performed using Western Blot Analysis. Results: Astrocyte-conditioned medium pretreatment of neurons showed both an increase in spinal cord neuron viability and a decrease in LDH release in a dose-dependent pattern. Moreover, pretreatment seems to attenuate glutamate-induced apoptotic cell death, antagonise glutamate-induced up-regulation of Caspase-3 expression and downregulate Bcl-2/Bax protein expression ratio. Conclusions: By attenuating glutamate-induced apoptotic cell death in primary cultured spinal cord neurons of rats, ACM seems to provide a neuroprotective effect by regulating apoptosis-related protein expression. Our results provide an experimental basis for clinical applications and potential therapeutic use of ACM in SCI.
引用
收藏
页码:803 / 808
页数:6
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