磷酸化DARPP-32在戊四氮诱导大鼠癫痫模型中的表达及其作用的研究  

Research on Expression and Function of Phosphorylated DARPP-32 on Pentylenetetrazol-induced Epilepsy Model of Rat

作　　者：王伟文[1] 廖晓阳[2] 杨正辉[1] 林航[1] 王庆松[1] 吴俞宪 刘榆[1] 

机构地区：[1]成都军区总医院神经内科,成都610083 [2]四川大学华西医院全科医学科/金卡病房,成都610041

出　　处：《生物医学工程学杂志》2014年第3期637-641,共5页Journal of Biomedical Engineering

基　　金：四川省科技厅应用基础研究发展计划基金资助项目(2008JY0071)

摘　　要：本文应用免疫酶组织化学及免疫荧光双标技术,将30只健康雄性SD大鼠分为一个对照组和5个实验组[即癫痫持续状态(SE)发作后1、6、24、48和72h组],研究磷酸化DARPP-32(p-DARPP-32)在PTZ诱导的癫痫发作大鼠前脑区域的表达(动态变化)和分布,并计算p-DARPP-32阳性表的神经元细胞数,以及与非磷酸化DARPP-32在大鼠前脑神经元的共存情况。结果表明,PTZ诱导大鼠SE发作1h和6h时,p-DARPP-32在大鼠前脑神经元的表达达到高峰,24h后开始逐渐下降;p-DARPP-32在大鼠前脑皮层、海马及纹状体神经元的胞质、胞核中有明显表达。免疫荧光双标记显示,p-DARPP-32与DARPP-32在上述区域出现高比例的共存。本研究提示p-DARPP-32在癫痫发作中可能起着重要作用。The present study is to explore the change process and distribution of phosphorylated DARPP-32 （p- DARPP-32） in rat brain including cortex, hippocampus and striatum and to further deduce whether p-DARPP-32 was possibly involved in epilepsy induced by repetitive low doses of pentylenetetrazol （PTZ）, PTZ-induced epilepsy model in rat was established with 30 male SD rats randomly divided into 6 groups, control group and five trial groups [PTZ 1 h,PTZ 6 h,PTZ 24 h,PTZ 48 h and PTZ 72 h respectively, after onset of status epilepticus （SE）]. Immunohisto- chemistry and immunofluorescence double-labeling were used to detect the temporal time change and distribution of p-DARPP-32 expression and to analyze the eoexpression of DARPP-32 and p-DARPP-32 in rat brain after the onset of PTZ-induced generalized SE. The results showed that there was a temporal time change of p-DARPP-32 expres- sion in rat brain after the onset of SE. The number of p-DARPP-32-positive cells increased significantly and reached the peaks at the ends of 1 hour and 6 hours after the onset of SE, but decreased at the end of 24 hours. The moderate to strong p-DARPP-32-immunopositive neurons were observed in cortex, hippocampus and striatum, and located in cell cytoplasm and cell nucleus. Further immunofluorescence double-labeling revealed that denser colocalization of p- DARPP-32 and DARPP-32 in the neurons existed in the area mentioned above. Therefore, PTZ-induced SE may cause phosphorylation of DARPP-32 in rat brain. The temporal time change and distribution of p-DARPP-32 suggest that phosphorylation of DARPP-32 may be involved in PTZ-induced epilepsy in rat brain including cortex, hippocam- pus and striatum, and p-DARPP-32 may play a central role in the onset of SE.

关 键 词：磷酸化DARPP-32 癫痫 戊四氮 大鼠 

分 类 号：R742.1[医药卫生—神经病学与精神病学]