摘要
目的观察慢病毒介导的神经元特异性生长抑素(somatostatin,SST)过表达对大鼠海马电点燃癫痫的抑制作用。方法成年Sprague—Dawley(sD)大鼠随机分为假手术组(Sham)、癫痫模型组(EP)、空载慢病毒注射癫痫模型组(Lenti—pSyn—EGFP,LV—EGFP):海马齿状回(dentategyms,DG)部位注射LV—EGFP病毒5μl和SST重组慢病毒注射癫痫模型组(Lenti—pSyn—SST-2A.EGFP,LV—SST):在海马齿状回、内嗅皮质(medial entorhinal codex,MEC)或杏仁核(amygdala,Amy)注射LV.SST病毒5μl;采用海马电点燃刺激法建立大鼠癫痫模型,观察各组大鼠行为学变化,免疫组化检测大鼠海马和大脑皮质内SST、神经元核蛋白(NeuN)以及微管相关蛋白2(MAP2)的表达变化。结果LV.SST(DG)组、LV—SST(MEC)组及LV—SST(Amy)组首次达到V级癫痫发作电流阈值分别为[(143.8±3.8)μA、(142.5±4.1)μA,(142.5±5.3)μA],与癫痫组[(136.3±5.3)μA]相比有所增加,且前4轮刺激达到V级的电流值均明显升高(P〈0.05);癫痫组和SST重组慢病毒注射组(LV—SST)大鼠电点燃后检测发现,在CA1、CA3和DG部位的SST和NeuN阳性表达细胞数均低于Sham组(P〈0.05),其中LV—SST组SST和NeuN表达降低幅度较癫痫组两者降低幅度小(P〈0.05),同时与癫痫组相比,LV—SST组大鼠MAP2的阳性表达较多。结论慢病毒介导的SST过表达对大鼠海马电点燃癫痫发作具有一定的抑制作用,对海马区神经元具有保护作用,为颞叶癫痫基因治疗提供新思路。
Objective To explore the effect of lentiviral vector-mediated somatostatin (SST) ex- pression on seizures induced by hippoeampal kindling in rats. Methods Adult Spragne-Dawley rats were randomly divided into the sham group (Sham), epilepsy group (EP), Lenti-pSyn-EGFP (LV-EGFP) group and Lenti-pSyn-SST-2A-EGFP (LV-SST) group.The rats in LV-EGFP group were subsequently electrically hippocampal kindled and LV-EGFP (5 μl) was injected into dentate gyrus (DG). The rats in LV-SST group were kindled and LV-SST (5 μl) was injected into the dentate gyms (DG), medial entorhinal cortex (MEC) or amygdala (Amy). Seizure severity was evaluated and immunohistochemical staining was em- ployed to detect the expression of SST, neuron specific nuclear protein (NeuN) and microtubule associated protein 2 (MAP2). Results The current values to the first stage V seizure of LV-SST (DG) group, LV-SST (MEC) group or LV-SST (Amy) group ( (143.8±3.8) μA, ( 142.5±4.1 ) μA, (142.5±5.3) μA,respeetive- ly) were significantly increased compared with that of epilepsy (EP) group ((136.3±5.3) μA) , and V stage current values of LV-SST groups in each stimulation day were higher than that of EP group except the fifth stimulation day (P〈0.05). After kindling, SST expression and NeuN-positive neurons of EP group and LV- SST groups were less than that of Sham group in CA1 ,CA3 and DG. SST and NeuN neurons loss in LV-SST groups were less than that of EP group (P〈 0.05 ) and groups were higher than that in EP group. Conclusion MA192 immunohistochemistry stainings in LV-SST Lentiviral vector-mediated somatostatin expression suppresses seizures and can rescue the neuronal damage of seizure induced by kindling in hippocampus, which may provide a new method of gene therapy for temporal lobe epilepsy.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2017年第7期594-599,共6页
Chinese Journal of Behavioral Medicine and Brain Science
基金
基金项目:江苏省教育厅科研项目(14KJB310021)
江苏省普通高校研究生创新计划项目(KY-LX15_1474)
关键词
癫痫
生长抑素
海马
慢病毒
基因治疗
Epilepsy
Somatostatin
Hippocampus
Lentivirus
Gene therapy