神经元特异性烯醇化酶在神经元氧糖剥夺损伤模型复氧后的表达变化  

Changes of the Expression of Neuron Specific Enolase After the Reoxygenation of Neuron Model Damaged by Oxygen-glucose Deprivation

作　　者：徐伟华[1] 赵冬[2] 戴晶[2] 刘祺[2] 许晖[2] 黄啸元 王业忠[2] 

机构地区：[1]石河子大学医学院,新疆石河子市832000 [2]石河子大学医学院第一附属医院神经外科

出　　处：《中国全科医学》2015年第20期2444-2447,共4页Chinese General Practice

基　　金：国家自然科学基金资助项目(81360185);新疆生产兵团博士基金资助项目(2011BB016)

摘　　要：目的探讨神经元特异性烯醇化酶(NSE)在体外培养神经元氧糖剥夺损伤模型复氧后的水平变化。方法体外原代培养24 h内的新生SD大鼠海马区神经元,应用免疫荧光染色及电子显微镜鉴定神经元,应用体外氧糖剥夺建立大鼠海马区神经元缺血低氧损伤模型,分别对复氧1、6、12、24、48、72 h神经元提取蛋白质,Western blotting法检测不同时间点神经元损伤模型中NSE表达水平。结果培养第4天的细胞免疫荧光染色显示神经元细胞核染色清晰,形态典型,突起走形如网状,阳性率为(93.6±1.6)%。各组NSE蛋白表达比较,差异有统计学意义(F=500.75,P<0.01)。实验组损伤后各时间点NSE蛋白表达高于对照组,细胞培养24、48 h NSE蛋白表达均高于其他时间点(P<0.05)。结论 NSE可作为自发性蛛网膜下腔出血(SAH)后早期脑损伤(EBI)标志物,能够反映脑组织的损伤程度。Objective To explore the changes of the expression of neuron specific enolase（NSE） after the reoxygenation of in vitro culture neuron model damaged by oxygen-glucose deprivation.Methods In vitro primary culture was conducted on the neurons in hippocampus area of newborn SD rats within 24 hours.Evaluation of the neurons was carried out by using immunofluorescent staining and electron microscope.By oxygen-glucose deprivation,the damaged neuron model which was ischemic and hypoxic was established.Protein was extracted from the model 1,6,12,24,48 and 72 hours after reoxygenation,and Western blotting method was used to test NSE expression level in the model at different time points.Results On day 4 during in vitro culture,immunofluorescent staining showed clear nuclear staining,typical shape of net with protuberance and deformation and a positive rate of（93.6 ± 1.6） %.The two groups were significantly different in NSE protein expression level（F = 500.75,P 0.01）.The trial group was higher（P 0.05） than the control group in NSE protein expression at different time points after neuron damage.The NSE protein expression levels at hour 24 and hour 48 during in vitro culture were higher（P 0.05） than those at other time points.Conclusion NSE could be used as an indicator of SAH and EBI,for it could reflect the damage degree of brain tissue.

关 键 词：蛛网膜下腔出血 原代细胞培养 海马 神经元 氧糖剥夺 神经元特异性烯醇化酶 

分 类 号：R743.35[医药卫生—神经病学与精神病学]