摘要
目的 观察N 甲基 D 门冬氨酸受体 (NMDAR ,NR)主亚基 (NR1)M3M4环重组肽自身抗血清对不同浓度谷氨酸诱导的神经元死亡的保护作用。方法 用NR1M3M4环重组肽免疫Balb/C(H 2 d)小鼠制备抗血清 ,用锥虫蓝染色法和原位末端标记 (TUNEL)法观察抗血清对原代培养海马神经元兴奋毒性坏死和凋亡的保护效应。结果 在高浓度谷氨酸 (5 0 0 μmol/L)条件下 ,阳性抗血清可以使神经元坏死减少 16 % ;在低浓度谷氨酸 (5 0 μmol/L)作用下 ,阳性抗血清保护可使神经元凋亡率减少 13%。结论 NMDAR重组肽自身免疫抗血清具有抗兴奋毒性作用 ,这一结论为免疫防治兴奋毒性脑损伤策略的建立提供了坚实的体外基础。
Objective To investigate whether the humoral autoimmune response targeting the NR1 subunit of NMDA receptor has neuroprotective activity or not, we observed the protection of serum against M3-M4 loop of NR1 from neuroexcitotoxicity induced by various concentration of glutamate in vitro. Methods The model of glutamate excitotoxicity on primarily cultured hippocampal neurons from new born Wistar rats were set up. Vaccination with M3-M4 loop recombinant peptide of NR1 was prepared as antiserum from Balb/c mice(H-2d). Protective effects of serum on glutamate-induced necrosis and apoptosis of hippocampal neurons were observed using Trypan blue dye staining and terminal deoxynucleotidyl transferase-mediated biotimylated deoxyuridine triphosphate nickel end labeling (TUNEL) method. Results At a high concentration of glutamate (500 μmol/L), the positive antiserum was found protecting 16% of neurons from necrosis, and at low concentration of glutamate (50 μmol/L), the positive antiserum had protection of 13% of neurons from apoptosis. Conclusion Serum against M3-M4 loop of NR1 might protect the cultured hippocampal neurons from excitotoxicity, suggesting the humoral autoimmune response targeting the NR1 subunit of NMDA receptor might have neuroprotective activity.
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2003年第5期366-369,共4页
Chinese Journal of Neurology
基金
国家自然科学基金 ( 3 0 0 70 2 67)
中国博士后科学基金 ( 2 0 0 1)
辽宁省重点科技攻关课题 ( 2 0 0 12 2 60 0 5 )
