NMDA受体在匹罗卡品癫痫小鼠海马星形胶质细胞活化中的作用-东南大学学报医学版

NMDA受体在匹罗卡品癫痫小鼠海马星形胶质细胞活化中的作用

单位：东南大学医学院, 江苏南京 210009

分类号：R-33;R742.1

出版年·卷·期（页码）：2017·36·第二期（129-136）

摘要：

目的：探讨在匹罗卡品诱导的小鼠癫痫持续状态（SE）模型中，N-甲基-D-天门冬氨酸（NMDA）受体在海马星形胶质细胞增生中的作用及其可能的分子机制。方法：将32只6周龄雄性C57/BL6小鼠随机分为对照组、SE组、SE后注射NMDA受体拮抗剂MK-801组和单纯注射MK-801组，每组8只。腹腔注射300 mg·kg^-1匹罗卡品建立SE模型。免疫组化方法检测各组小鼠海马脑区星形胶质细胞的形态差异，同时利用western Blot方法检测海马脑区神经胶质纤维酸性蛋白（GFAP）的表达和转录因子cAMP反应元件结合蛋白（CREB）的磷酸化水平。结果：SE组小鼠海马星形胶质细胞出现显著活化现象，免疫组化结果显示SE组小鼠海马GFAP免疫反应强度显著高于对照组（P<0.01），而在SE后给予MK-801阻断NMDA受体则显著抑制了SE诱导的海马星形胶质细胞活化。与免疫组化结果一致，Western Blot结果显示SE组小鼠海马GFAP蛋白表达水平显著高于对照组（P<0.05），而在SE后给予MK-801阻断NMDA受体则显著抑制了SE诱导的海马GFAP蛋白表达水平的升高。同时我们的研究结果发现，SE组小鼠海马CREB蛋白磷酸化水平显著高于对照组，而阻断NMDA受体则显著抑制了SE诱导的海马CREB蛋白磷酸化。结论：匹罗卡品癫痫小鼠持续状态7 d后海马星形胶质细胞被显著活化，而这一活化过程依赖于NMDA受体的激活。同时，海马星形胶质细胞活化的过程伴随CREB的磷酸化，提示海马CREB磷酸化参与了NMDA受体介导的匹罗卡品癫痫小鼠星形胶质细胞活化过程。

Objective: To study the effect of N-methy-D-asparate(NMDA) receptor on hippocampal astrocyte activation in pilocarpine-induced epileptic mice and the possible mechanism involved. Methods: Thirty two C57/BL6 mice were randomly divided into control group, status epilepticu(SE) group, NMDA receptor antagonist MK-801 treated SE group and MK-801 alone treated group, with 8 mice in each group. SE epilepsy model was established by intraperitoneal injection of 300 mg·kg^-1 pilocarpine. Immunohistochemistry was employed to determine hippocampal astrocyte activation in different group of mice. Western Blot was used to detect hippocampal glial fibrillary acidic protein(GFAP) content and transcription factor cAMP response element binding protein(CREB) phosphorylation level. Results: Hippocampal astrocytes were largely activated in SE mice. Immunohistochemistry results showed that GFAP immunostaining intensities in the hippocampus of SE mice were significantly increased as compared to those of control mice. NMDA receptor antagonist MK-801 treatment after SE, however, suppressed SE-induced hippocampal astrocyte activation. Consistent with the immunohistochemistry results, our Western Blot showed that GFAP level in SE mice was significantly higher than that of control mice. MK-801 treatment after SE, however, suppressed SE-induced increase of GFAP level. Meanwhile, our Western Blot result showed that phosphorylation level of cAMP response element binding protein(CREB) was significantly increased in SE mice as compared to that of control mice. Blocking NMDA receptor by MK-801, however, suppressed the SE-induced increase of CREB phosphorylation. Conclusion: Pilocarpine-induced SE can activate hippocampal astrocytes, which are dependent on the activation of NMDA receptors. Meanwhile, hippocampal astrocyte activation in pilocarpine-induced SE mice is accompanied with CREB phosphorylation, suggesting CREB phosphorylation is involved in NMDA receptor-mediated hippocampal astrocytes activation in pilocarpine induced epileptic mice.

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