神经肽Y对海人酸致痫大鼠中NR1、NR2A受体的影响

Influence of NR1 and NR2A receptors of neuropeptide Y in kainic acid-kindled rat

目的探讨重组腺相关病毒载体神经肽Y(r AAV2/1-NPY-EGF-P)干预前后海人酸致痫大鼠海马中NR1、NR2A表达的变化。方法将36只雄性Wistar大鼠随机等分为对照组、海人酸致痫大鼠组(KA组)以及神经肽Y(NPY)干预组(NPY组)各12只。KA组和NPY组均在海马CA3区注射海人酸建立为慢性癫痫模型,造模成功后,NPY组向脑室注射10l滴度为5×10^(11)μg·mL^(-1)的r AAV2/1-NPYEGFP进行基因转染,KA组不注射病毒,对照组海马CA3区注射等量的生理盐水。各组动物分别于基因转染后2w、4w,取海马保存于液氮。每组动物选取12只,每个时间点6只大鼠,用免疫组化方法检测各组NR1,NR2A的表达情况。对试验结果采用双盲法进行统计,应用半定量积分的方法,把每个时间点的大鼠CA3区NPY的阳性细胞率和阳性细胞着色强度进行分级记分,最后依据两项之积计算其阳性强度(IHS)。结果 2w及4w时KA组和NPY组NR1、NR2A阳性细胞数量、染色强度和IHS分数均高于对照组,(P<0.05);4w时NPY组阳性细胞比例、染色强度和IHS评分均低于KA组,(P<0.05)。结论 NR1、NR2A受体的低表达可能参与了NPY抑制癫痫发作的发病机制。

Objective To explore the expression change of NR1and NR2A in hippocampal of the kainic acid-kindled rats intervened by recombinant adeno-associated virus vector neuropeptide Y(rAAV2/1-NPY-EGFP).Method36cases of male Wistar rats were divided into control group,kainic acid-kindled rats group(KA group)and neuropeptide Y intervention group(NPY group).KA and NPY group were made into chronic epilepsy model by injecting kainic acid in hippocampal CA3region.KA group wasn't given virus,though NPY group was injected10lrAAV2/1-NPY-EGFP(5×1011μg·ml-1)in ventricle.The control group was injected normal saline of same volume.The hippocampal organizations were obtained2and4weeks after gene transfection.12cases of rats were selected ineach group,and6cases for each time.The expressions of NR1and NR2A were tested through immunohistochemical method.The test results were counted by double blind method,and the positive intensity in hippocampal CA3region was calculated by semi quantitative integral method.Results The positive cell number of NR1and NR2A,staining intensity and IHS fraction in KA and NPY group were higher than those of control group2and4weeks after gene transfection(P<0.05).The positive cell number,staining intensity and IHS fraction in NPY group were lower than those of KA group4weeks after gene transfection(P<0.05).Conclusion The low expression of NR1and NR2A receptors may be involved in the pathogenesis of NPY inhibition of seizures.