摘要
目的观察大鼠癫痫持续状态（SE）后72 h内海马组织中NKCC1及GABAARαl表达的动态变化,以及使用anti-BDNF干预后其表达的变化,探讨NKCC1与GABAARαl在SE中的可能机制及其之间的关系,以及BDNF在SE发生过程中对NKCC1和GABAARαl表达的影响。方法健康成年雄性SD大鼠90只,随机分为生理盐水对照组（10只）、SE组（40只）和干预组（40只）。制作Li Cl-PILO癫痫模型（SE组）和造模前3~4 h侧脑室注射anti-BDNF（干预组）,选取SE后2、6、24和72 h为时间点。观察大鼠在SE诱发过程中的行为学差异、海马NKCC1及GABAARαl蛋白的表达改变、海马各区NKCC1及GABAARαl蛋白的时空分布等特点。结果干预组SE诱发成功率较SE组低,SE诱发成功后生存状态好、死亡率低。SE组NKCC1蛋白在SE后均高于对照组,干预组NKCC1蛋白表达在2 h显著增加,6 h达最高,24 h后开始明显恢复,72 h时接近正常。两组在海马不同亚区,SE后2 h开始,NKCC1在CΑ1及CΑ3区均显著增高。SE组GABAARαl蛋白在SE后2~6 h表达显著下调,24 h降至最低,至72 h突然增高。干预组GΑBAARαl蛋白表达在SE后2 h达最低值,6 h后上调,72 h达最高值,SE组与干预组GABAARαl在CA1、CA3区的表达呈进行性下调,但干预组变化幅度较小。结论 NKCC1蛋白表达在SE后的上调可能是SE后GABAAR介导的兴奋性作用产生的重要机制之一;Αnti-BDNF干预可通过抑制BDNF的表达,延缓癫痫发生并减弱癫痫发作的严重程度。
【Objective】To observe the expressions of Na＋-K＋-2Cl-cotransporter 1（NKCC1） and GΑBΑΑR αl proteins in rat hippocampus 72 hours after status epilepsy（SE） and their changes after using sheep brain-derived neurotrophic factor（BDNF） neutralization antiserum（anti-BDNF）, so as to investigate the role of NKCC1 in the formation of SE, the relationship between NKCC1 and GΑBΑΑRαl, and the effect of BDNF on modulating the activation and expressions of NKCC1 and GΑBΑΑRαl in SE. 【Methods】A total of 90 SD male rats were randomly divided into NS control group（10 rats）, SE group（40 rats） and experiment group（40 rats）. The rats of the SE group received Li Cl-PILO i.p for induction of SE. The rats of the experiment group received injection of anti-BDNF 3 ~4 h before the induction of SE. The rats of both SE group and experiment group were randomly subdivided into 4 subgroups（10rats in each subgroup）, and were sacrificed 2, 6, 24 and 72 h after SE induction. The changes of ethology of the rats during and after SE induction were observed and evaluated. The expression and distribution of NKCC1 and GΑBΑΑRαl in hippocampus were detected with immunohistochemistry and Western Blot. 【Results】The successful rate of SE induction and the mortality rate in the experiment group were lower than those in the SE group. The expression of NKCC1 was substantially up-regulated in the SE group compared to the NS control group. In the experiment group the expression of NKCC1 obviously increased at 2 h, reached the peak at 6 h, started to decrease at 24 h and nearly returned to normal at 72 h after SE ended. Up-regulation of NKCC1 was significant in CA1 and CA3 areas after SE. The GΑBΑΑRαl protein expression in the SE group was markedly down-regulated 2 ~6 h after SE,reached the lowest level at 24 h, but greatly up-regulated at 72 h after SE. The GΑBΑΑRαl expression in the experiment group was down-regulated and reached the lowest level at 2 h, but up-regulated from 6 h to 72 h. The G
出处
《中国现代医学杂志》
CAS
北大核心
2015年第18期1-7,共7页
China Journal of Modern Medicine
基金
湖南省科技厅项目(No:2012FJ4367)
湖南省科技厅项目(No:2012FJ3041)
