氯胺酮诱导大鼠原代培养皮层神经元凋亡机制研究  

A Mechanism Study on Apoptosis of Primary Cultured Cortical Neurons Induced by Ketamine in Rats

作　　者：李建立[1] 于洋[2] 吴红海[2] 侯艳宁[2] 

机构地区：[1]河北省人民医院麻醉科,石家庄050051 [2]解放军白求恩国际和平医院药剂科,石家庄050082

出　　处：《解放军医药杂志》2016年第7期21-23,28,共4页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：河北省卫生厅指令性课题(ZL20140095);2015年政府资助临床医学优秀人才培养和基础课题研究项目(361003-6)

摘　　要：目的研究氯胺酮诱导大鼠原代培养皮层神经元凋亡的机制。方法原代培养7 d的大鼠皮层神经元随机分为溶剂对照组和氯胺酮组。氨胺酮组给予氯胺酮终浓度为100μmol/L,溶剂对照组给予相同容积的生理盐水,两组分别处理24 h后,用光学显微镜观察两组神经元形态学变化,流式细胞仪检测神经元的凋亡率,罗丹明染色检测线粒体膜电位的变化,蛋白免疫印迹法检测神经元cyt-c蛋白水平。结果与溶剂对照组比较,氯胺酮组光镜下神经元数量明显减少,胞体立体感消失,细胞轮廓不清,神经元轴突断裂;神经元凋亡率和cyt-c水平增加,线粒体膜电位下降(P<0.01)。结论氯胺酮可诱导发育期原代培养皮层神经元凋亡,线粒体膜电位下降以及cyt-c的释放可能是凋亡发生的主要原因。Objective To investigate mechanism of apoptosis of primary cultured cortical neurons induced by Ketamine in rats. Methods Cortical neurons were primarily cultured for 7 d, and then were divided into control, group which was treated with equal volume of physiological saline, and Ketamine group, which was treated with 100μmol/L fi-nal concentration of Ketamine. After treatment for 24 h, morphological changes of neurons were observed using micro-scope;apoptosis rates were detected using flow cytometer;potential changes of mitochondrial membrane were measured u-sing fluorescent dye Rhodamine 123 （Rh123）;Western blot method was used to detect cyt-c neurons level. Results Compared with those in control group, neurons number significantly decreased, three-dimensional sense of cell bodies dis-appeared, and uncleared outline and axonotmesis were found under microscope observation in Ketamine group;values of apoptosis rate and cyt-c neurons level increased （P〈0. 01）, which mitochondrial membrane potential decreased in Ket-amine group （P〈0. 01）. Conclusion Ketamine induced apoptosis of primary cultured cortical neurons, decreasing lev-el of mitochondrial membrane potential and releasing cyt-c may be important cause of apoptosis incidence.

关 键 词：氯胺酮 原代细胞培养 皮层 神经元 凋亡 大鼠 Sprague-Dawley 

分 类 号：R971.2[医药卫生—药品] R-332[医药卫生—药学]