钠氢交换体1在氯化锂-匹罗卡品点燃癫痫大鼠模型海马组织内的表达

Expression of NHE1 in hippocampus of epileptic rat model kindled by lithium pilocarpine

目的 检测钠氢交换体1(NHE1)在氯化锂-匹罗卡品点燃癫痫大鼠模型海马组织中的表达。方法 选取雄性SD大鼠90只,其中45只采用腹腔注射氯化锂-匹罗卡品构建癫痫模型作为模型组,45只采用腹腔注射生理盐水组作为对照组;取模型组和对照组大鼠各36只,采用蛋白印迹(Western blot)法检测造模后第1、3、7、14、30及60天时2组大鼠海马组织匀浆中NHE1蛋白的定量表达;取模型组和对照组大鼠各3只,采用免疫组织化学分析造模后第14天时2组大鼠海马组织NHE1的蛋白表达;取模型组和对照组大鼠各6只,采用逆转录聚合酶链式反应(RT-PCR)检测造模后第14天2组大鼠海马组织NHE1 m RNA的的表达特点。结果造模后第1、3、7、14、30及60天时,模型组大鼠海马组织匀浆中NHE1蛋白表达较对照组增高(P<0.05),且模型组大鼠海马组织NHE1蛋白表达从第1天开始增高、第14天时达到高峰、第30天和第60天逐渐下降(P<0.05);造模后第14天时,模型组大鼠海马组织不同亚区NHE1阳性表达较对照组相应区域增强,且NHE1m RNA水平也高于对照组(P<0.05)。结论 氯化锂-匹罗卡品点燃模型癫痫大鼠海马组织中NHE1表达随时间呈动态变化,且在第14天表达最高,提示NHE1可能与癫痫的发生发展过程密切相关。

Objective To explore the expression of Na+/H+ exchanger-1(NHE1) in the hippocampus of epileptic rat model kindled by lithium-pilocarpine.Methods A total of 90 male SD rats were included in the present study.The epileptic model group was established by intraperitoneal injection with lithium pilocarpine(n=45).The control group was done by intraperitoneal injection with normal saline(n=45).Thirty six rats were selected from model and control group respectively,which expression of NHE1 protein in the hippocampus was detected by western blot on the 1st,3rd,7th,14th,30th,and 60th day.Three rats were selected from model and control respectively,which expression of NHE1 protein in hippocampus was detected by immunohistochemistry on the 14th day after modeling.Six rats were selected from model group and control group respectively,which expression of NHE1 mRNA in hippocampus was detected by reverse transcription polymerase chain reaction(RTPC R) on the 14th day after modeling.Results The expression of NHE1 protein in hippocampus of model group was higher than that in the control group(P<0.05) on 1st,3rd,7th,14th,30th,and 60thday after modeling.The expression of NHE1 protein in model group kept increasing from the 1st day and reached the peak on the 14 th day,but decreased gradually on the 30th and 60th day(P<0.05).On the 14 th day after modeling,the positive expression of NHE1 in different sub-regions of hippocampus of model group was more obvious than that in the control group.The mRNA level of NHE1 in the model group was higher than that in the control group(P<0.05).Conclusion The expression of NHE1 in the hippocampus of rats kindled by lithium pilocarpine shows a dynamic change with time,and the most intensive expression is on the 14th day,which suggested that NHE1 might closely relate to the occurrence and development of epilepsy.