中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (23): 2059-2065.doi: 10.4103/1673-5374.147932

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

谷氨酸受体6羧基端过表达可抑制海人藻酸诱导大鼠海马神经元的损伤

  

  • 收稿日期:2014-09-22 出版日期:2014-12-10 发布日期:2014-12-10
  • 基金资助:

    国家自然科学基金(30800309, 81372172)

Overexpression of C-terminal fragment of glutamate receptor 6 prevents neuronal injury in kainate-induced seizure via disassembly of GluR6-PSD95-MLK3 signaling module

Jie Mou 1, Xiaomei Liu 2, Dongsheng Pei 3   

  1. 1 Jiangsu Key Laboratory of Targeted Drug and Clinical Application, Xuzhou Medical College, Xuzhou, Jiangsu Province, China
    2 School of Basic Medical Science, Xuzhou Medical College, Xuzhou, Jiangsu Province, China
    3 Jiangsu Key Laboratory of Biological Cancer Therapy, Xuzhou Medical College, Xuzhou, Jiangsu Province, China
  • Received:2014-09-22 Online:2014-12-10 Published:2014-12-10
  • Contact: Dongsheng Pei, Ph.D., Jiangsu Key Laboratory of Biological Cancer Therapy, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China,dspei@xzmc.edu.cn.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 30800309, 81372172; the Educational Science Foundation of Jiangsu Province, China, No. 10KJB350005; the Xuzhou Science Foundation in China, No. XZZD1153; the President Special Grant of Xuzhou Medical College in China, No. 09KJZ20; and a grant from the Zhenxing Project Foundation of  XZMC.

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摘要：

课题组以往研究发现小肽Tat-GluR6-9c (含有谷氨酸受体6 羧基末端的9个氨基酸的融合肽)可以进入海马神经元，在脑缺血模型中通过谷氨酸受体6•PSD-95•MLK3信号模块可抑制MLK3和JNK的激活，在海人藻酸诱导的癫痫模型中发现谷氨酸受体6•PSD-95•MLK3信号模块在海马CA1区组装。为此，实验观察了腺病毒转载的谷氨酸受体6羧基端能否抑制谷氨酸受体6•PSD-95•MLK3信号模块的组装并减少海马CA1区海人藻酸诱导神经元细胞的死亡。 将海人藻酸腹腔注射于SD大鼠诱导癫痫模型，免疫沉淀和免疫印迹的方法检测在海人藻酸注射后不同时间点海马CA1区谷氨酸受体6•PSD-95信号模块的组装，发现谷氨酸受体6过表达抑制海人藻酸注射6 h谷氨酸受体6和PSD-95的相互作用，并阻止海人藻酸诱导的大鼠海马神经元中JNK, MLK3和MKK7的激活，而免疫组织化学染色显示，注射谷氨酸受体6后能显著减少海人藻酸引起的神经元细胞死亡和凋亡，说明谷氨酸受体6的羧基端过表达可抑制海人藻酸诱导大鼠海马神经元的损伤。

关键词: 神经再生, 脑损伤, 海马神经元损伤, 癫痫, 腺病毒, GluR6, PSD95, MLK3, 海人藻酸, 细胞凋亡, JNK, 国家自然科学基金

Abstract:

Our previous study showed that when glutamate receptor (GluR)6 C terminus-containing peptide conjugated with the human immunodeficiency virus Tat protein (GluR6)-9c is delivered into hippocampal neurons in a brain ischemic model, the activation of mixed lineage kinase 3 (MLK3) and c-Jun NH2-terminal kinase (JNK) is inhibited via GluR6-postsynaptic density protein 95 (PSD95). In the present study, we investigated whether the recombinant adenovirus (Ad) carrying GluR6c could suppress the assembly of the GluR6-PSD95-MLK3 signaling module and decrease neuronal cell death induced by kainate in hippocampal CA1 subregion. A seizure model in Sprague-Dawley rats was induced by intraperitoneal injections of kainate. The effect of Ad-Glur6-9c on the phosphorylation of JNK, MLK3 and mitogen-activated kinase kinase 7 (MKK7) was observed with western immunoblots and immunohistochemistry. Our findings revealed that overexpression of GluR6c inhibited the interaction of GluR6 with PSD95 and prevented the kainate-induced activation of JNK, MLK3 and MKK7. Furthermore, kainate-mediated neuronal cell death was significantly suppressed by GluR6c. Taken together, GluR6 may play a pivotal role in neuronal cell death.

Key words: nerve regeneration, brain injury, hippocampal neuronal injury, seizures, adenovirus, GluR6, PSD95, MLK3, kainate, apoptosis, JNK, NSFC grants, neural regeneration