Nectin-1在锂-匹罗卡品癫痫大鼠海马中的表达

Expression of nectin-1 in the hippocampus of LICL-PILO induced epileptic rats

目的为探讨Nectin-1在癫痫形成中的作用,我们采用经典的氯化锂-匹罗卡品大鼠癫痫模型,在点燃持续状态后不同时间点观察Nectin-1蛋白在大鼠脑海马的表达变化规律。方法①选用正常成年雄性SD大鼠,随机分为正常对照组(n=10)与模型组(n=50),模型组采用氯化锂-匹罗卡品腹腔注射法,对照组用等量生理盐水腹腔注射。成功点燃后存活的大鼠以(3 d、1周、2周、4周、6周)为研究时间点随机分为5个亚组,每组10只。②用Western-blot、免疫组化等经典方法观察Nectin-1蛋白在癫痫大鼠模型中的表达变化及规律。结果实验组大鼠腹腔注射匹罗卡品后,经过急性发作期及潜伏期、慢性期,出现癫痫反复的自发性发作,对照组未见急性及慢性发作。免疫组化染色对照组Nectin-1蛋白主要在胞膜及胞浆表达,在海马CA3区深染区域集中于透明层(SL)。癫痫组2周、4周时Nectin-1在齿状回有较多的阳性细胞,主要表现为胞浆和核膜棕色,并向内分子层分布。Western-blot与对照组相比,在匹罗卡品诱导的SE之后2周表达升高,一直持续到慢性期4周、6周。结论Nectin-1蛋白在癫痫大鼠模型SE后慢性期海马组织中表达较正常增高,可能与癫痫的形成机制有关。

Objective To explore possible roles of nectin -1 in the epileptogenesis, we investigated theexpression pattern of nectin-1 in the hippocampus of lithium-pilocarpine-kindled rat. Methods Sixty healthy adultmale Sprague-Dawley rats were randomly divided into seizure group（n=50） and control group（n=10）. Seizure and controlwas injected with the pilocarpine （50 mg/kg） and saline of the same volume, respectively. Rats survived after Statusepilepticus（SE） were then divided into5 subgroups （3d,1w,2w,4w,6 w） and each group consisted of 10 rats. Theexpression profile of Nectin -1 was detected by using immunohistochemistry and western blot. ResultImmunohistochemistry showed that the Stratum granulosum （SG） of Dentate gyrus （DG） within hippocampus waspositively stained and the Nectin-1 stains were almost located at the membrane of DG neurons. In rats of 2 and4weeks after status epilepticus （SE）, stains in the DG were more concentrated compared with the control group andextended to the inner molecular layer （IML）.Western blot analysis showed that nectin-1 expression was upreguelatedat 2, 4 and6 weeks after SE compared with the control group（P〈0.05）. Conclusion The present study has detecteda high expression of nectin-1 in the hippocampus of LICL-PILO-kindled rats, indicating an involvement of nectin-1in epileptogenesis.