神经肽Y对海人酸致痫大鼠中GABA_B受体的影响

Effect of neuropeptide GABA_B on Y receptor in rats with epilepsy induced by sea

目的探讨重组腺相关病毒载体神经肽Y(rAAV2/1-NPY-EGF-P)干预前后海人酸致痫大鼠海马中GABABR表达的变化。方法将36只雄性Wistar大鼠随机分为三组:KA组(海人酸致痫大鼠组)、NPY组(神经肽Y干预组)及NS组(对照组)。KA组大鼠海马CA3区注射海人酸,完成慢性模型,造模成功后不注射病毒;NPY组大鼠海马CA3区注射海人酸,完成慢性模型,造模成功后向脑室注射10μL滴度为5×1011 vg/mL的神经肽Y进行基因转染;NS组海马CA3区注射等量的0.9%NaCl。三组大鼠各12只,在基因转染后2周、4周分别取6只观察其发作行为学变化,并采用RT-PCR检测GABABR的表达,以GAPDH为标准参照基因,以之定量分析基因表达的RQ值(相对定量值)。结果通过RT-PCR发现,KA组、NPY组及NS组、海马组织均有GABABR表达。KA组GABABR的基因表达较NS组呈下降趋势(P<0.05),NPY组GABABR的基因表达呈上升趋势,差异有统计学意义(P<0.05)。结论 KA组中GABABR的表达明显下降,NPY组中GABABR的表达呈上升趋势。说明GABABR基因表达的降低可能是导致癫痫发生、发展的重要因素。NPY可能通过改变抑制性神经递质的GABABR的表达来达到调控癫痫发作的目的。

Objective To explore the mechanism of epilepsy by observing the alternation of GABA receptors （GABABR） in hippocampus before and after neuropeptide Y （rAAV2/1 NPY-EGFP） intervention in epileptic rats induced by kainic acid. Methods 36 male Wistar rats were randomly divided into three groups （12 for each group）, which were KA group（Kainic Acid Group） ,NPY group （Neuropeptide Y Group） , and NS Group （Control group）. KA group was by injecting kainic acid into the hippocampal CA3 region of the rats without injecting virus vector. NPY group was by injecting kainic acid into the hippocampal CA3 region of the rats and injected 10 μL. rAAV2/1- NPY-EGFP into the intraventricular, with a titer of 5 ）〈 10^11 vg / mL. NS group was by injecting equal volume saline （0.9% NaC1） in hippocampal CA3 region. Rats were selected to observe epilepsy occurrence after transfection two and four weeks （six rats for each）. Then RT-PCR was performed to quantify the expression of GABAB receptor, u- sing GAPDH as reference genes to get the target gene RQ value （the relative quantitative expression values）. Re- suits KA group no significant changes after the intervention, NPY group attack reduced significantly after the inter vention. KA group and NPY group hippocampus were GABAeR expression by RT-PCR. KA group of GABABR gene expression than the NS group decreased （P〈0.05）. NPY group of GABABR gene expression than the KA group were increased and there were statistical significantly differences （P〈0.05）. GABABR gene expression chan- ges was little small at two time-points. Conclusion GABABR gene may lead to epilepsy, an important factor in devel- opment. NPY may achieve the purpose of control seizures by changing the inhibitorythe neurotransmitter GABABR expression.