摘要
目的 探讨神经肽Y基因（rAAV2/1-hNPY-EGFP）转染对大鼠癫痫发作行为、脑电图（electroencephalogram,EEG）及其海马磷酸化Tau蛋白的影响.方法 将实验组动物分为3组：对照组、KA组及NPY组,每组24只.NPY组向侧脑室注射10μl的rAAV2/1-NPY-EGFP（滴度为5×1011v.g./ml）,KA组向侧脑室注射等量生理盐水.对照组在海马CA3区及侧脑室注射等量生理盐水.利用视频监视系统观察注射后2周和4周各组大鼠癫痫发作情况、发作潜伏期以及EEG,同时应用Western blotting方法检测大鼠海马中磷酸化Tau蛋白的表达.结果 （1）2周时NPY组大鼠行为学发作级别[（12.13±8.06）分]与KA组相比[（12.10±8.07）分]无明显差异（P＞0.05）,4周时NPY组大鼠行为学发作级别降低[（6.06±3.78）分]、发作潜伏期延长[（79.06± 8.83） min]、脑电图癫痫波放电频率减少、波幅降低（P＜0.05）,对照组无发作；（2）与对照组相比,KA组和NPY组大鼠在2周和4周时磷酸化Tau蛋白表达水平均明显增加（P＜0.05）；与KA组相比（1.87±0.23 RQ value）,4周时NPY组大鼠海马组织中磷酸化Tau蛋白表达水平（1.15±0.16 RQ value）显著降低（P＜0.05）.结论 磷酸化Tau蛋白在癫痫大鼠海马组织中的表达变化与癫痫发作密切相关；rAAV2/1-hNPY-EGFP基因转染可以明显减轻大鼠癫痫发作级别,并可延长发作潜伏期.rAAV2/1-hNPY-EGFP基因转染可能通过抑制KA诱导的大鼠癫痫发作时磷酸化Tau蛋白的表达而发挥抗癫痫作用.
Objective To discuss the effect of gene transfection of rAAV2/1-NPY-EGFP on KA-induced rat seizures,EEG and the expression of hippocampal phosphorylated Tau protein.Methods Altogether 72 healthy male Wistar adult rats were randomly divided into three groups：control group,KA group and NPY group（n=24）.The epileptic models were established by the injection of KA 2 μl （0.4 μg/μl） five times to the right side of the hippocampus CA3 area every three days.rAAV2/1-NPY-EGFP group,in which 10 μl of rAAV2/1-NPY-EGFP （titer 5× 1011 v.g./ml） was injected to the lateral ventricle in successful rats chronic model,while KA group was injected with an equal dose of saline.The control group was injected with an equal dose of saline both in the hippocampal CA3 area and the lateral ventricle.The seizure situation,the onset latency and EEG were observed at 2 weeks and 4 weeks after vector injection.Then the expression of phosphorylated Tau protein in hippocampus were detected with Western blotting.Results （1） Scale and latency of each seizure onset in rats of rAAV2/1-NPY-EGFP group （12.13 ± 8.06） had no significant difference at 2 weeks （P＞ 0.05） compared with KA group （12.10± 8.07）.The scale of seizure in rats of rAAV2/1-NPY-EGFP group（6.06±3.78） significantly reduced at 4 weeks（P ＜0.05）.Latency of seizure onset （79.06±8.83min） significantly increased at 4 weeks（P＜0.05）,EEG epileptic discharge frequency and wave amplitude decreased （P＜ 0.05） at 4 weeks.The control group had no seizures.（2）Compared with the control group,the expression of phosphorylated Tau protein in KA group and NPY group significantly increased（P＜0.05） at 2 weeks and 4 weeks,and the expression of phosphorylated Tau protein in the NPY group （1.15±0.16 RQ value） at 4 weeks significantly decreased （P＜0.05） compared with the KA group（1.87± 0.23 RQ value）.Conclusion rAAV2/1-NPY-EGFP gene transfection significantly reduces scale of seizure onset and prolongs latency of seizure onset in KA-induced rat model.rAAV2/1-NPY-EGFP gene transfection may play anti-epileptic and neuroprotective effects through inhibiting the expression of phosphorylated Tau protein in hippocampus of KA-induced epileptic rat model.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2014年第2期104-106,共3页
Chinese Journal of Behavioral Medicine and Brain Science
基金
河北省卫生厅科研基金项目（20110209）
关键词
神经肽Y
癫痫
基因转染
磷酸化TAU蛋白
Neuropeptide Y
Epilepsy
Gene transfection
Phosphorylated Tau protein
