环磷酸腺苷反应元件结合蛋白调控耐药性癫痫大鼠海马内脑源性神经营养因子的表达  

Regulatory effects of cAMP response element-binding protein on hippocampal brain-derived neurotrophic factor level in a rat model of drug-resistant epilepsy

作　　者：李婧萱 时黛 伍国锋 Li Jingxuan;Shi Dai;Wu Guofeng(Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学,贵州省贵阳市550004 [2]贵州医科大学附属医院,贵州省贵阳市550004

出　　处：《中国组织工程研究》2023年第35期5659-5664,共6页Chinese Journal of Tissue Engineering Research

基　　金：贵州医科大学附属医院国家自然科学基金(NSFC)面上基金培育计划项目(gyfynsfc-2021-8),项目负责人:伍国锋。

摘　　要：背景:耐药性癫痫的形成机制尚不清楚,其中环磷酸腺苷反应元件结合蛋白(cAMP response element-binding protein,CREB)及脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)在耐药性癫痫形成过程中的作用已被广泛研究。目的:使用慢病毒转染法调控正常大鼠海马CREB表达水平,探究其对耐药性癫痫形成过程及海马内BDNF表达的影响。方法:使用过表达慢病毒或干扰慢病毒调控大鼠海马中CREB表达水平,检测病毒转染结果;大鼠注射病毒7 d后构建氯化锂-匹罗卡品癫痫模型,随后使用苯妥英钠和苯巴比妥进行耐药性筛选,筛选出耐药性癫痫大鼠,分为过表达组、过表达对照组、干扰组、干扰对照组,观察筛药过程中大鼠癫痫发作次数变化,使用蛋白质免疫印迹法及实时荧光定量PCR法检测大鼠海马中CREB及BDNF表达水平。结果与结论:①病毒转染后14 d,大鼠海马区满布绿色荧光,病毒转染水平较好;②药筛前过表达组大鼠癫痫发作次数多于过表达对照组,干扰组大鼠癫痫发作次数少于干扰对照组;药筛后过表达组大鼠癫痫发作次数多于过表达对照组,干扰组大鼠癫痫发作次数少于干扰对照组;③过表达组大鼠CREB、BDNF表达水平高于过表达对照组,同样干扰组大鼠CREB、BDNF水平低于干扰对照组;④结果表明,调控大鼠海马内CREB表达水平会影响大鼠癫痫发作及耐药性癫痫的形成,通过调控CREB表达可能会对耐药性颞叶癫痫大鼠海马内BDNF表达水平产生影响。BACKGROUND:The formation mechanism of drug-resistant epilepsy is still unclear,and the role of cAMP response element-binding protein(CREB)and(brainderived neurotrophic factor(BDNF)in the formation of drug-resistant epilepsy has been widely studied.OBJECTIVE:To regulate the expression of CREB in the hippocampus of normal rats by lentivirus transfection and to explore its effect on the formation of drugresistant epilepsy and the expression of BDNF in the hippocampus of rats.METHODS:Lentivirus overexpression or interfering was used to regulate the expression level of CREB in the hippocampus of rats and detect the virus transfection results.Lithium-pilocarpine induced epilepsy models were established in rats 7 days after lentivirus injection.Sodium phenytoin and phenobarbital were then used for resistance selection.Drug-resistant epileptic rats were screened and divided into overexpression group,overexpression control group,interference group and interference control group.The frequency of epileptic seizures during the screening process was observed and the expression levels of CREB and BDNF in the hippocampus of rats were measured by western blot and real-time fluorescence quantitative PCR.RESULTS AND CONCLUSION:Fourteen days after virus transfection,the hippocampus of rats was covered with green fluorescence,indicating that the virus transfection level was good.The frequency of epileptic seizures in the overexpression group was higher than that in the overexpression control group and the frequency of epileptic seizures in the interference group was lower than that in the interference control group.The expression levels of CREB and BDNF in the overexpression group were higher than those in the overexpression control group,while the expression levels of CREB and BDNF in the interference group were lower than those in the interference control group.These results indicate that the regulation of CREB expression level in the hippocampus of rats can affect epileptic seizures and formation of drug-resistant epilepsy in rat

关 键 词：耐药性癫痫 脑源性神经营养因子 环磷酸腺苷反应元件结合蛋白 CREB-BDNF-TrkB信号通路 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R363.21