N-甲基-D-门冬氨酸受体主亚基单克隆抗体抗谷氨酸兴奋毒性的体外实验研究  

Glutamate-neuroexcitotoxicity protective study of monoclonal antibody against human NMDA receptor key subunit

作　　者：康晓楠[1] 孙长凯[1] 范明[2] 丁爱石[2] 赵杰[1] 王吉庆[3] 韩大跃[3] 施广霞[4] 

机构地区：[1]大连医科大学脑疾病研究所,116027 [2]军事医学科学院基础医学研究所 [3]解放军第二一○医院 [4]大连医科大学病理生理学教研室

出　　处：《中华医学杂志》2003年第18期1603-1606,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目 (3 0 0 70 2 67);中国博士后科学基金资助项目(2 0 0 1);辽宁省重点科技攻关课题 (2 0 0 12 2 60 0 5 )

摘　　要：目的　研究人N 甲基 D 门冬氨酸受体 (NMDAR ,NR)主亚基 (NR1)单克隆抗体mAbN1是否对兴奋毒性脑损害有防护作用。方法　取 10只新生大鼠海马神经细胞进行体外原代培养 ,建立了谷氨酸对培养海马神经细胞兴奋毒性神经损伤模型 ,观察抗体保护后神经细胞形态 ,检测细胞存活率 (锥虫蓝拒染法 )和细胞培养上清乳酸脱氢酶漏出量 ,每组 2个平行皿 ,实验重复 4次。结果　经 0 .3μmol/LmAbN1预处理后 ,神经元具有明显抗谷氨酸兴奋毒性损伤能力 ,存活率提高 35 % ;抗体表位合成肽可以阻断这种保护 ,提示该抗体的保护功能是特异性的。结论　NR1单克隆抗体mAbN1具有体外抗兴奋毒作用 。Objetive To study if monoclonal antibody against human NMDA receptor key subunit (NR1) may protect neurons from excitotoxicity. Methods We cultured primary hippocampal neurons from 10 newborn rats and made the glutamate excitotoxicity model, examined the ratio of surviving neuron (Trypan blue dye staining ) and LDH assay to study the protective effects of mAbN1,There were 2 plates in parallel per group, the experiments were repeated 4 times. Results 0.3 μmol/L mAbN1 could protect cultured hippocampal neurons from neuroexcitotoxicity induced by 500 μmol/L glutamate with increased survival rate 35%. Epitope peptide could block the protection of mAbN1, which suggests that the protection of mAbN1 is specific. Conclusion mAbN1 is a novel NMDAR blocker with in vitro neuroprotective activity which may provide a basis for antibody therapy of neuroexcitotoxicity.

关 键 词：N-甲基-D-门冬氨酸受体 主亚基 单克隆抗体 抗谷氨酸兴奋毒性 体外实验 细胞培养 缺血性脑损伤 

分 类 号：R741[医药卫生—神经病学与精神病学]