《中国康复理论与实践》

《中国康复理论与实践》

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磷酸化环磷腺苷反应元件结合蛋白对持续惊厥后海马细胞凋亡调控基因表达的影响①

苑爱云，刘秋燕，李小平，侯梅   

  1. 青岛大学医学院附属妇女儿童医院神经康复科，山东青岛市266034。
  • 出版日期:2016-02-25 发布日期:2016-06-01

Effect of Phosphorylation Cyclic Adenosine Monophosphate Response Element Binding Protein on Expression of Apoptotic Regulated Genes in Hippocampus after Status Convulsivus

YUAN Ai-yun, LIU Qiu-yan, LI Xiao-ping, HOU Mei   

  1. Department of Neurology Rehabilitation, Women and Children's Hospital, Medical School, Qingdao University, Qingdao, Shandong 266034, China
  • Published:2016-02-25 Online:2016-06-01

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摘要： 目的观察外源性磷酸环磷腺苷反应元件结合蛋白(pCREB)抗体对惊厥持续状态(SC)后海马细胞凋亡调控基因Bcl-2和c-Jun 表达的影响，探讨pCREB 在惊厥性脑损伤中的作用及其调控机制。方法成年Wistar 鼠48 只随机分为正常对照组(NC组，n=24)和惊厥持续状态组(SC 组, n=24)。SC组采用氯化锂-匹罗卡品腹腔注射诱发SC模型。两组均根据脑室注射内容分为无注射组、生理盐水注射组(NS组)和抗pCREB注射组(抗pCREB组)3 个亚组，每个亚组8 只。注射后6 h 处死动物。取双侧海马采用免疫组化和原位杂交(ISH)观察Bcl-2 蛋白/mRNA、c-Jun 蛋白/mRNA的分布，并进行半定量分析。结果NC组中，3 个亚组注射侧Bcl-2 蛋白/mRNA表达均无显著性差异(P>0.05)，SC组中抗pCREB组低于无注射组和NS组(P<0.05)。NC组和SC组中，3 个亚组注射侧c-Jun 蛋白/mRNA 表达均有非常高度显著性差异(P<0.001)，NS 组和抗pCREB 组c-Jun 蛋白/mRNA 表达高于无注射组(P<0.05)，抗pCREB组高于NS组(P<0.05)。结论SC后脑室内注射外源性抗pCREB抗体可下调SC后海马Bcl-2 蛋白/mRNA表达，上调c-Jun蛋白/mRNA表达。

关键词: 持续惊厥, 磷酸化环磷腺苷反应元件结合蛋白, Bcl-2, c-Jun, 大鼠

Abstract: Objective To explore the effect of the exogenous phosphorylation cyclic adenosine monophosphate response element binding protein (pCREB) antibody on the expression of apoptotic regulated genes (Bcl-2 and c- Jun) in hippocampus after status convulsivus (SC), to elucidate the role and regulation mechanism of pCREB in convulsive brain injury. Methods Seizures were induced in 24 adult Wistar rats with lithium-pilocarpine intraperitoneal injection (SC group), another 24 rats were as the normal controls (NC group). Each group was divided into no injection subgroup, normal saline injection subgroup and anti pCREB subgroup according to the injection contents of lateral ventricle, with 8 cases in each group. They were sacrificed 6 hours after injection. Both the protein and mRNA expression of Bcl-2 and c-Jun in bilateral hippocampus were detected by immunohistochemistry and in situ hybridization, respectively. Results There was no significant difference in Bcl-2 protein/mRNA expression among 3 subgroups in the NC group (P>0.05). In the SC group, the expression of Bcl-2 protein/mRNA were lower in the anti pCREB subgroup than in the no injection subgroup and normal saline injection subgroup (P<0.05). There was significant difference in c-Jun protein/mRNA expression among 3 subgroups in both NC group and SC group (P<0.001). The expression of c-Jun protein/mRNA was higher in the normal saline injection subgroup and the anti pCREB subgroup than in the no injection group (P<0.05), especially in the anti pCREB subgroup (P<0.05). Conclusion Exogenous anti-pCREB antibody can down-regulate the expression of Bcl-2 and up-regulate the expression of c-Jun in hippocampal cells after SC.

Key words: status convulsivus, phosphorylation cyclic adenosine monophosphate response element binding protein, Bcl-2, c-Jun, rats