摘要
目的 研究氯化锂-匹罗卡品致癫痫持续状态后大鼠海马区钾离子通道Kv1.6的表达及分布变化,探讨钾离子通道Kv1.6与癫痫发作的关系.方法 48只健康雄性Sprague-Dawley大鼠按配对比较法随机分为实验组和对照组,每组按随机区组法继续随机分为6h、1d、2d和3d4个观察时间点亚组（n=6）.通过大鼠脑电监测记录大鼠脑电变化情况,通过尼氏染色观察脑组织病理改变,采用免疫组织化学染色和Western印迹方法检测各时间点大鼠海马区Kv1.6的表达及分布变化.结果 （1）脑电监测：正常大鼠脑电图表现为波幅较均匀一致的0波,痫性发作后开始出现慢波、棘波,波幅、节律不规则,癫痫持续状态过程中表现为长程的棘波活动.（2）尼氏染色：癫痫持续状态后6h未发现明显形态学及神经元数量改变;癫痫持续状态后1d,海马区神经元结构松散,神经元数量减少;癫痫持续状态后2、3d,海马区神经元进一步减少,且出现神经元肿胀、变形、尼氏小体减少甚至消失.（3）蛋白表达检测：癫痫持续状态后6h,Kv1.6在海马CA3和CA1区表达较对照组无明显变化（均P＞0.05）;癫痫持续状态后1、2d及3d,Kv1.6在海马CA3区（0.117 ±0.040、0.135±0.029、0.112±0.050）表达较对照组（0.192±0.054、0.201±0.062、0.184±0.042）显著减少（t=3.861、3.745、5.579,均P＜0.05）,在CA1区（0.114±0.023、0.082±0.031、0.106 ±0.043）表达较对照组（0.165±0.036、0.187±0.025、0.172±0.052）也显著降低（t=3.221、6.360、3.645,均P＜0.05）.癫痫持续状态后6h、1d、2d及3d,Kv1.6在海马DG区表达较对照组无明显变化（P＞0.05）.结论 Kv1.6的表达下调可能与癫痫的发作相关.
Objective To investigate the expression and distribution of Kv1.6 in rat hippocampus after status epilepticus （SE） induced by lithium and pilocarpine to explore the relationship between Kv1.6 and epileptogenesis.Methods Forty-eight healthy male Sprague-Dawley rats were randomly divided into experiment group and control group.The rats in each group were randomly assigned into 6-hour,1-day,2-day and 3-day subgroups （n =6）.Electroencephalography （EEG） of rats in both groups were recorded to show the electrical activity of the rat brains.Nissel staining was performed to investigate the pathological changes,and immunohistochemistry and Western blotting were applied to analyze the expression and distribution of Kv1.6.Results （1） The EEG of rats in control group showed α wave with uniform amplitude,while the EEG in rats subjected to pilocarpine injection showed slow waves and spike waves with irregular amplitude and rhythm.Sustained spike waves were observed during SE.（2） Nissl staining showed that 6 hours after SE,the neural morphology of hippocampi in experiment group was as the same to that in control group.There was no change in neuronal morphology and numbers.However,1 day after SE,the structure of hippocampi was discrete and the number of neurons was decreased.Two and 3 days after SE,the number of neurons in hippocampi decreased further.Swelling and deformation of neurons were seen.Nissl bodies emerged,decreased or even disappeared.（3） There was no statistically significant difference in the expression of Kv1.6 in CA3 and CA1 areas between experiment group and control group 6 hours after SE （all P〉0.05）.One day,2 days and 3 days post SE,the expression of Kv1.6 in CA3 area （0.117 ±0.040,0.135 ±0.029,0.112 ±0.050） decreased obviously compared to control group （0.192±0.054,0.201 ± 0.062,0.184 ±0.042;t---3.861,3.745,5.579,all P 〈0.05）;and the expression of Kv1.6 in CA1 area （0.114 ± 0.02,0.082 ± 0.031,0.106 ± 0.043） decreased significantly compared to control group （0.165±0.036,0.187 ±0.025,0.172 ±0.052;t=3.221,6.360,3.645,all P〈0.05）.The expression of Kv1.6 did not show statistically significant difference in DG area between experiment group and control group at all time points examined （P 〉 0.05）.The Western blotting showed the same trend.Conclusion The decreased expression of Kv1.6 may play a role in the epileptogenesis.
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2016年第9期709-714,共6页
Chinese Journal of Neurology
基金
国家自然科学基金资助项目（81300985）
辽宁省自然科学基金资助项目（2015020549）
大连市卫生计生委计划项目
