自发性癫痫大鼠海马电压门控性钠通道β_1亚基表达与基因突变分析  

The β_1 subunit expression and gene mutation analysis of voltage-gated sodium channel in spontaneously epileptic rat hippocampus

作　　者：郭凤[1] 高青华[2] 封瑞[1] 孟秀君[3] 孙凤[1] 俞军龄[1] 毛小元[1] 谢妮[1] 刘睿[1] 吴坤灿[1] 蔡际群[1] 

机构地区：[1]中国医科大学药学院药物毒理学教研室,辽宁沈阳110001 [2]中国医科大学生理学教研室,辽宁沈阳110001 [3]中国医科大学附属第四医院心内科,辽宁沈阳110001

出　　处：《中国药理学通报》2009年第11期1434-1437,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30270535)

摘　　要：目的与正常Wistar大鼠作对比,探讨自发性癫痫大鼠(SER)海马中电压门控性钠通道β1亚基mRNA与蛋白的表达情况及基因突变分析。方法应用RT-PCR技术检测β1亚基mRNA的表达;免疫组化方法定位β1亚基蛋白;免疫印迹方法检测β1亚基蛋白的表达;直接测序技术筛查β1亚基全部外显子基因突变位点。结果SER海马β1亚基mRNA表达高于Wistar大鼠(P<0.05);免疫组化结果显示β1亚基蛋白在SER大鼠海马各区中均有表达,且位于细胞膜上;β1亚基蛋白的表达高于Wistar大鼠(P<0.01);直接测序法筛查了β1亚基全部外显子的突变位点,结果未见任何突变。结论SER海马β1亚基表达异常可能会促进癫痫样兴奋活动的产生,进而构成SER癫痫表型的发作基础。Aim To detect voltage-gated sodium channel （VGSC） β1 subunit expression and mutation analysis in spontaneously epileptic rat （SER） hippocampus compared with that of control Wistar rats.Methods The mRNA expression of β1 subunit was detected by RT-PCR;the protein distribution of β1 subunit was detected by immunohistochemistry;the protein expression of β1 subunit was detected by Western blot;the β1 subunit mutation analysis of five exons was detected by DNA sequencing.Results The mRNA expression of β1 subunit in SER hippocampus was significantly higher than that in control rat hippocampus by RT-PCR（P0.05）;the protein distribution of β1 subunit in SER hippocampus was detected in all the areas of hippocampus by immunohistochemistry;the protein expression of β1 subunit was significantly increased by Western blot（P0.01）. However,there was no sequence mutation for five exons of β1 subunit by means of mutation analysis.Conclusion Sodium channel β1 subunit abnormal expression at the mRNA and protein levels of SER hippocampus might result in the epileptiform activity and underlie the observed seizure phenotype in SERs.

关 键 词：自发性癫痫大鼠 电压门控性钠通道 亚基 基因突变 癫痫 外显子 

分 类 号：R-332[医药卫生] R322.81