大鼠皮质及海马神经元体外缺氧缺糖模型建立  

Establishment of oxygen-glucose deprivation model of rats′ cortex and hippocampal neurons in vitro

作　　者：郑敏[1] 郭莲军[2] 吕青[2] 何治[2] 徐旭林[2] 喻欣[2] 宗贤刚[2] 

机构地区：[1]咸宁学院药理教研室 [2]华中科技大学同济医学院药理系,湖北武汉430030

出　　处：《中国老年学杂志》2006年第3期333-335,共3页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(No.30371639)

摘　　要：目的建立体外培养大鼠皮质及海马神经元缺氧缺糖模型。方法取培养10—12d的皮质及海马神经元，用古连二亚硫酸钠1mmol／L的无糖Earle液取代正常含血清培养基，分别培养2、4、8、12、24及36h后，MTr法检测细胞活力，并测定培养液中LDH活力。结果连二亚硫酸钠1mmol／L合并基质缺糖2—36h，大鼠皮质及海马神经元细胞活力均显著降低（P〈0．01），并呈时间依赖性下降，至培养36h时细胞存活率分别仅为25．4％和24．2％；培养液LDH活力则均显著升高（P〈0．01）。并呈时间依赖性上升；且皮质及海马神经元培养液LDH活力变化与其细胞活力变化均呈高度负相关（r=-0．983，-0．992，P〈0.01）。结论连二亚硫酸钠合并基质缺糖可建立体外培养大鼠皮质及海马神经元缺氧缺糖模型。Objective To establish oxygen-glucose deprivation model of rats＇ cortex and hippocampusal neurons in vitro. Methods The cortex and hippocampul neurons cultured for 10 - 12 days were exposed to Earle＇s solution with sodium dithionite（ Na2S2O4 ） 1 mmol/L and free glucose in stand of original culture medium containing serum for 2,4,8,12,24 and 36 h respectively. 3-（ 4,5-dimethyhhiazol-2yl ）- 2,5-diphenyherazolium bromide（MTT） bssay was applied to evaluate the neuronal vigor. Biochemical method was used to determine lactate dehydrogenase（LDH） activity. Results The rat cortex and hippocampal neurons vigor significantly decreased in a time-dependent manner cultured by Earle＇s solution with sodium dithionite 1 mmol/L and free glucose for 2 - 36 h（ P 〈 0. 01 ）, and the neuron survival rate decreased to 25.4% and 24. 2% respectively 36 h later. However, the LDH activity significantly increased in a time-depondent manner（ P 〈 0. 01 ） showing great negative correlation to neurons vigor （ r = - 0. 983, - 0. 992, P 〈 0. 01 ）. Conclusions The oxygen-glucose deprivation model of rats＇ cortex and hippocampal neurons in vitro could be established successfully by Earle＇s solution with sodium dithionite 1 mmol/L and free glucose.

关 键 词：缺氧缺糖 皮质 海马 模型 

分 类 号：R743.9[医药卫生—神经病学与精神病学]