调控RET依赖性GDNF信号通路对大鼠癫痫模型的影响  

The Effect of RET-dependent GDNF Signaling-regulation in a Rat Model of Epilepsy

作　　者：田沁涛 黑悦 伊西才[1] 魏礼洲[1] 刘卫平[1] 

机构地区：[1]第四军医大学附属西京医院神经外科,陕西西安710032

出　　处：《现代生物医学进展》2017年第19期3628-3631,共4页Progress in Modern Biomedicine

基　　金：陕西省科技统筹基金资助项目(2013KTCL03-08)

摘　　要：目的:探究双侧海马CA1区立体定向注射anti-GDNF抗体对匹鲁卡品诱导的大鼠癫痫模型的影响。方法:选择成年雄性SD大鼠60只,并随机分为3组,即假手术组(sham组,n=20)、癫痫模型组(model组,n=20)和GDNF抑制剂组(anti-GDNF组,n=20)。使用氯化锂-匹鲁卡品腹腔注射诱导癫痫模型,sham组只给予氯化锂,anti-GDNF组在造模前2 h给予大鼠双侧海马CA1区立体定向注射anti-GDNF抗体。在造模后1、3、7 d观察大鼠癫痫的发作频率,7 d后采用脑电图监测(EEG)测定脑电波的变化情况,通过免疫组化方法测定海马CA1区域神经元数量变化(Neu N表达水平),造模后1 d时使用western blot方法测定海马CA1区GDNF、RET和P53蛋白的表达。结果:Model组大鼠棘-慢波数量明显高于Sham组,anti-GDNF组以上指标较model组显著减少(P<0.05);Model组海马CA1区神经元大量凋亡,但anti-GDNF组凋亡较model组显著减少(P<0.05)。与Sham组比较,在癫痫发作后1 d,model组的GDNF、RET表达水平上调,P53表达水平下降(P<0.05),而anti-GDNF组大鼠海马CA1区GDNF、RET表达较model组明显下调,P53表达水平显著上降(P<0.05)。结论:双侧海马CA1区立体定向注射anti-GDNF抗体能够减少癫痫发作,并对海马神经元起到保护作用,可能与其抑制GDNF/RET/P53信号通路有关。Objective：To investigate the effect of bilateral stereotactic administration of anti-GDNF antibody in the hippocampal CA1 region of a rat model of pilocarpine-induced epilepsy.Methods：60 adult male SD rats were randomly divided into the sham group （n=20）,model group （n=20） and anti-GDNF group （n=20）.Lithium chloride and pilocarpine were intraperitoneally injected to induce seizures.The sham group was given lithium chloride only and anti-GDNF group was given a stereotactic injection of anti-GDNF antibody into bilateral hippocampal CA1 region 2h before seizure induction.Then video monitoring of seizure incidence of the epileptic rats was done at 1,3,7d and EEG monitoring was performed at 7 d.Immunohistochemical staining was used to analyze the amount of neurons （NeuN expression） in the hippocampal CA1 region at 7 d,and western blot was carried out to explore the expression of protein levels of GDNF,RET and P53 in the hippocampal CA1 region at 1 d.Results：Model group tend to have more spike-slow waves than Sham group,and anti-GDNF group tend to have less than Model group in the parameters above （P〈0.05）;the amount of apoptotic neurons in the hippocampal CA1 region of Model group were large,but apoptotic cells of the anti-GDNF group were comparatively less than Model group （P〈0.05）;compared to Sham group,protein levels of GDNF and RET were up-regulated and P53 down-regulated in Model group 1 d after seizure induction （P〈0.05）,however the protein levels of GDNF and RET were obviously down-regulated and P53 up-regulated in the anti-GDNF group compared to Model group （P〈0.05）.Conclusions：Bilateral stereotactic administration of anti-GDNF antibody in the hippocampal CA1 region could reduce the incidence of seizure and have protective effects on the hippocampal neurons,which was probably related to suppression of GDNF/RET/P53 signaling.

关 键 词：癫痫 转染重排基因 神经胶质细胞源性的神经营养因子 神经元保护 脑电图 

分 类 号：R-33[医药卫生] R742.1