摘要
目的观察G蛋白耦联受体激酶相关蛋白1(GIT1)在氯化锂-匹罗卡品致模型大鼠海马组织中的表达变化,以探讨GIT1在癫发生发展过程中的作用机制。方法共42只无特定病原体级成年雄性Wistar大鼠,制备氯化锂-匹罗卡品致模型,随机分为对照组和癫组(癫发作后1、3、7、14、30、60 d),荧光定量聚合酶链反应、Western blotting法和免疫组织化学染色检测各观察时间点大鼠海马组织GIT1表达变化。结果癫组大鼠GIT1 m RNA自癫持续状态后急性期第1和3天即升高(P=0.012,0.002),至潜伏期第7和14天持续升高(P=0.003,0.001),至慢性期第30和60天达峰值水平(均P=0.000);GIT1蛋白自急性期即升高,慢性期持续升高,但与对照组相比,差异未达统计学意义(均P>0.05),至慢性期达峰值水平(均P=0.000)。至慢性期第30天时,癫组大鼠海马CA1区、齿状回和海马旁皮质GIT1蛋白表达水平均高于对照组(P=0.000)。结论癫大鼠海马组织GIT1表达上调,可能通过调节细胞骨架动态重组而影响兴奋性神经网络,从而参与癫的发生。
Objective To investigate the expression changes of G-protein-coupled receptor kinase-interacting protein 1（GIT1） in lithium-pilocarpine-induced epileptic rat model and explore its role in thegenesis and development of epilepsy.MethodsThe lithium- pilocarpine- induced model of statusepilepticus（SE） was established in 42 specific pathogen free（SPF） male adult Wistar rats, and those ratswere randomly divided into control group and 6 epilepsy groups（1, 3, 7, 14, 30, 60 d after SE）. Theexpression of GIT1 m RNA was detected by fluroescent quantitative polymerase chain reaction（PCR）, whilethe expression of GIT1 protein was examined by Western blotting and immunohistochemistry was applied totest the expression of CA1 region, dentate gyrus and parahippocampal cortex in rat hippocampus at differenttime points.ResultsGIT1 m RNA level rised in acute phase on 1st and 3rd day after SE（P = 0.012,0.002）, then increased continously in latency on 7th and 14 th day（P = 0.003, 0.001）, and reached the peakin chronic phase on 30 th and 60 th day（P = 0.000, for all）. GIT1 protein expression rised in acute phaseand increased continously in chronic phase, but there was no significant difference compared with controlgroup（P 0.05, for all）. Then, it reached the peak in chronic phase（P = 0.000, for all）. Until the 30 th day,the GIT1 expression of CA1 region, dentate gyrus and parahippocampal cortex in the hippocampus of rats in6 epilepsy groups was significantly higher than that of control group（P = 0.000, for all）.ConclusionsTheup-regulated expression of GIT1 in the hippocampus of epileptic rat was probably involved in the formationprocess of chronic epilepsy by regulating cytoskeleton dynamic regrouping to influence excitatory neural networks.
出处
《中国现代神经疾病杂志》
CAS
2015年第6期475-480,共6页
Chinese Journal of Contemporary Neurology and Neurosurgery
基金
四川省宜宾市科技计划重点项目(项目编号:2013SF008)~~
关键词
GTP结合蛋白质类
受体
G-蛋白偶联
癫
海马
疾病模型
动物
GTP-binding proteins
Receptors, G-protein-coupled
Epilepsy
Hippocampus
Disease models, animal
