尼莫地平诱导小脑颗粒神经元凋亡及机制研究  

Nimodipine promotes apoptosis of cerebellar granule neurons

作　　者：何伟文[1,2] 王翦[1,2] 闵志群[3] 李明昌[1,2] 罗景燕[4] 袁忠民[1,2] 

机构地区：[1]广州医学院第二附属医院神经外科 [2]广州医学院神经科学研究所,广东省广州市5102602 [3]广州医学院第二附属医院分子实验中心,广东省广州市510260 [4]中山大学中山医学院药理教研室,广东省广州市510089

出　　处：《国际神经病学神经外科学杂志》2010年第2期114-118,共5页Journal of International Neurology and Neurosurgery

基　　金：广东省科技计划项目(73055);广州市医药卫生科技一般引导项目(2009-YB-154)

摘　　要：目的探讨尼莫地平能否诱导小脑颗粒神经元凋亡及可能机制。方法取Sprague-Dawley(SD)大鼠小脑颗粒神经元,体外培养7d,用含10μM尼莫地平培养基处理神经元24h,对照组为含1‰二甲亚砜(DMSO)培养基处理细胞。Hoechst33258染胞核检测凋亡率。Westernblot检测c-Fos和c-Jun蛋白的表达水平;腺病毒作载体过表达c-Fos和负显性c-Jun突变体阻断c-Jun功能,检测能否抑制尼莫地平诱导的神经元凋亡,免疫细胞化学方法检测腺病毒感染率。结果对照组神经元的凋亡率为(10±4)%,尼莫地平处理6、12、24h后神经元的凋亡率分别为(13±4)%、(28±5)%、(45±3)%。尼莫地平处理使c-Fos表达下调,但c-Jun表达水平上调。腺病毒Ad-c-Fos和Ad-c-JunDN(负显性c-Jun突变体)对神经元感染率为85%以上,过表达c-Fos或负显性c-Jun突变体使尼莫地平诱导的神经元凋亡率从(42±6)%减少到(28±6)%或(20±3)%。结论尼莫地平通过下调c-Fos和上调c-Jun表达诱导小脑颗粒神经元凋亡。Objective To investigate whether Nimodipine induces apoptosis of cerebellar granule neurons and the possible mechanisms.Methods Cerebellar granule neurons（CGNs）from Sprague-Dawley（SD）rats were cultured in vitro for 7 days and treated with media containing 10 μM Nimodipine for 24 hrs.Cells were cultured in media containing 1‰ dimethyl sulfoxide（DMSO）as a control.Neuronal nucleic were stained by Hoechst 33258 for apoptotic analysis.Western blot was performed to detect the expression of c-Fos and c-Jun.Adenovirus was used as a vector to overexpress c-Fos and dominant negative c-JunDN in CGNs.Imnunocytochemistry was performed to determine the infection rate of adenovirus.Results The apoptotic rates for control CGNs were（10±4）%,and the apoptotic rates for CGNs treated with Nimodipine for 6,12 or 24 hrs were（13±4）%,（28±5）% or（45±3）%,respectively.Nimodipine treatment resulted in a downregulation of c-Fos and an upregulation of c-Jun.The infection rate of adenovirus in CGNs was more than 85%,Adenovirus-delivered overexpression of c-Fos or dominant negative c-Jun mutant attenuated Nimodipine-induced CGN apoptosis from（42±6）% to（28±6）% or（20±3）%.Conclusions Nimodipine induced CGN apoptosis through downregulating c-Fos expression and upregulating c-Jun expression.

关 键 词：尼莫地平 小脑颗粒神经元 凋亡 C-FOS C-JUN 大鼠 

分 类 号：R96[医药卫生—药理学]