CAJ | 학술논문

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。
목적：탐토신경원특이성희순화매（ NSE）재체외배양신경원양당박탈손상모형복양후적수평변화。방법체외원대배양24 h내적신생SD대서해마구신경원，응용면역형광염색급전자현미경감정신경원，응용체외양당박탈건립대서해마구신경원결혈저양손상모형，분별대복양1、6、12、24、48、72 h신경원제취단백질， Western blotting법검측불동시간점신경원손상모형중NSE표체수평。결과배양제4천적세포면역형광염색현시신경원세포핵염색청석，형태전형，돌기주형여망상，양성솔위（93.6±1.6）%。각조NSE단백표체비교，차이유통계학의의（ F=500.75，P<0.01）。실험조손상후각시간점NSE단백표체고우대조조，세포배양24、48 h NSE단백표체균고우기타시간점（P<0.05）。결론 NSE가작위자발성주망막하강출혈（SAH）후조기뇌손상（EBI）표지물，능구반영뇌조직적손상정도。
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 <br> 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.