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

发布时间：2018-06-24 14:06

本文选题：N-甲基-D-天门冬氨酸受体 + 匹罗卡品　；参考：《东南大学学报(医学版)》2017年02期

【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c AMP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(P0.01),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(P0.05),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。
[Abstract]:Aim: to investigate the role of N-methyl-Daspartate (NMDA) receptor in hippocampal astrocyte proliferation and its possible molecular mechanism in pilocarpine induced epileptic status (SE) model in mice. Methods: Thirty-two 6-week-old male C57 / BL6 mice were randomly divided into control group (SE group) and NMDA receptor antagonist group (MK-801 group) and MK-801 group (8 mice in each group). SE model was established by intraperitoneal injection of pilocarpine (300 mg kg ~ (-1). The morphological differences of astrocytes in hippocampus of each group were detected by immunohistochemical method. Western blot was used to detect the expression of glial fibrillary acidic protein (GFAP) and phosphorylation of transcription factor c-AMP response element binding protein (CREB) in hippocampus. Results the astrocytes in the hippocampus of the mice in the group of 10% SE were activated significantly. The results of immunohistochemistry showed that the intensity of GFAP immunoreactivity in hippocampus of SE group was significantly higher than that of control group (P0.01), but blocking NMDA receptor by MK-801 after SE significantly inhibited the activation of hippocampal astrocytes induced by SE. The expression level of GFAP protein in hippocampus of SE group was significantly higher than that of control group (P0.05), while blocking NMDA receptor by MK-801 after SE significantly inhibited the increase of GFAP protein expression induced by SE. At the same time, we found that the phosphorylation level of CREB protein in hippocampus of SE group was significantly higher than that of control group, while blocking NMDA receptor significantly inhibited the phosphorylation of CREB protein induced by SE. Conclusion: hippocampal astrocytes were significantly activated in pilocarpine epileptic mice 7 days later, and the activation process was dependent on the activation of NMDA receptor. At the same time, the activation of hippocampal astrocytes was accompanied by the phosphorylation of CREB, which suggested that the phosphorylation of hippocampal CREB was involved in the activation of astrocytes mediated by NMDA receptor in pilocarpine epileptic mice.
【作者单位】：东南大学医学院;
【基金】：国家自然科学基金资助项目(81673413) 江苏省自然科学基金资助项目(BK20141335) 高等学校博士学科点专项科研基金新教师类资助项目(20130092120043) 国家本科生科研训练计划(SRTP)项目(201610286134)
【分类号】：R742.1

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