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目的探讨重组腺相关病毒介导人源性神经肽Y基因(rAAV2/1-hNPY-EGFP)转染对红藻氨酸(kainicacid,KA)诱导大鼠癫痫发作行为和脑电图的影响。方法 66只雄性Wistar大鼠分为点燃组(n=54)和对照组(n=12)。点燃组以大鼠右侧海马CA3区注射0.4μg/μlKA1.5μl5次造成慢性癫痫大鼠模型后,又随机分为KA组、增强型绿色荧光蛋白(EGFP)组和神经肽Y(neuropeptide,NPY)组(n=18)。EGFP组脑室注射rAAV2/1-EGFP10μl,滴度为5×1011/ml;NPY组注射rAAV2/1-hNPY-EGFP10μl,滴度为5×1011/ml。分别于注射后2、3、4周,再次右侧海马CA3区注射1.5μlKA诱导癫痫发作,观察各组大鼠的行为和脑电图改变。结果对照组无癫痫发作;在发作级别、发作潜伏期、脑电痫波密度和脑电波幅方面,2、3周时NPY组与KA组、EGFP组相比无明显差异(P>0.05);4周时NPY组与KA组、EGFP组相比有明显差异(P<0.05),表现为发作级别降低,发作潜伏期延长,脑电痫波密度和脑电波幅均减少。结论 rAAV2/1-hNPY-EGFP基因转染可以抑制KA诱导的大鼠癫痫发作和海马神经元放电。
Objective To investigate the effect of rAAV2 / 1-hNPY-EGFP transfection on the behavior and electroencephalogram of rats induced by kainic acid (KA) induced by recombinant adeno-associated virus. Methods Sixty-six male Wistar rats were divided into igniting group (n = 54) and control group (n = 12). The rats in the ignited group were randomly divided into KA group, EGFP group and neuropeptide Y (NPY) group ) Group (n = 18). The intracerebroventricular injection of rAAV2 / 1-EGFP in EGFP group had a titer of 5 × 1011 / ml and 10μl of rAAV2 / 1-hNPY-EGFP in the NPY group with a titer of 5 × 1011 / ml. At 2, 3, and 4 weeks after injection, once again, 1.5 μl of KA was injected into CA3 area of hippocampus to induce seizures. The changes of behavior and EEG of the rats in each group were observed. Results There was no seizure in the control group. There was no significant difference between the NPY group and the KA group and the EGFP group at seizure level, seizure latency, wave density of EEG and EEG amplitude at 2 and 3 weeks (P> 0.05). 4 Compared with KA group and EGFP group, there was a significant difference (P <0.05) in NPY group at week 1, showing a decrease in seizure level, prolonged seizure latency, and decreased wave density and amplitude of EEG wave. Conclusion Transfection of rAAV2 / 1-hNPY-EGFP gene can inhibit KA-induced seizure and hippocampal neuronal discharge.