大脑皮质神经元氧糖剥夺／复氧后NF-κB P50及c-Rel蛋白的表达  

Expression of NF-κB subunits P50 and c-Rel protein in primary cortical neurons after oxygen glucose deprivation/reoxygenation

作　　者：何兰英[1] 罗勇[1] 

机构地区：[1]重庆医科大学附属第一医院神经内科、重庆市神经病学重点实验室,重庆400016

出　　处：《第二军医大学学报》2008年第11期1333-1336,共4页Academic Journal of Second Military Medical University

基　　金：重庆市卫生局资助项目(渝卫科教[2003]61号,03-2-179)~~

摘　　要：目的:探讨Wistar大鼠大脑皮质神经元氧糖剥夺/复氧(OGD/R)后不同时间点NF-κB亚基P50及c-Rel蛋白表达情况。方法:体外培养原代大脑皮质神经元,采用免疫细胞化学鉴定神经元,建立氧糖剥夺/复氧模型,以正常组、OGD 4h组、OGD 4h/R2h组、OGD 4h/R6h组、OGD 4h/R12h组、OGD 4h/R24h组为实验组,采用免疫细胞化学、Western印迹检测NF-κB亚基P50及c-Rel蛋白表达。结果:(1)免疫细胞化学(神经元烯醇化酶相关抗原及β-tubulin 3)鉴定所培养的细胞为神经元;(2)免疫细胞化学、Western印迹结果显示OGD 4h组NF-κB P50蛋白表达开始增加明显高于正常组(P<0.05),OGD 4h/R2h组、OGD 4h/R6h组NF-κB P50蛋白表达持续性升高,并在复氧后6h达高峰(P<0.01),此后逐渐下降,至OGD 4h/R 12h时与正常组有显著差异(P<0.05),OGD 4h/R 24h NF-κB P50蛋白表达降低与正常组比较无统计学差异(P<0.05);(3)NF-κB c-Rel蛋白表达OGD 4h组与正常组无差异(P>0.05),OGD 4h/R 2h至OGD 4h/R 12h表达持续增加,并在复氧后12h达高峰(P<0.01),OGD4h/R24h仍未恢复到正常水平(P<0.05)。结论:大鼠大脑皮质神经元氧糖剥夺/复氧后激活了NF-κB P50及c-Rel蛋白的表达,并具有时间相关性。Objective：To investigate the expression of nuclear factor-κB （NF-κB） subunits P50 and c-Rel protein in primary cortical neurons of Wistar rats at different time points of oxygen glucose deprivation/reoxygenation（OGD/R）. Methods： The neurons dissociated from the cortex of the neonatal rats were primary cultured and were identified by immunocytochemistry. OGD/R model was established. The study was divided into 6 groups according to different processing methods,including normal group,OGD 4 h treated,OGD 4 h/R 2 h treated,OGD 4 h/R 6 h treated,OGD 4 h/R 12 h treated and OGD 4 h/R 24 h treated groups. The expression of NF-κB P50 and c-Rel protein in neurons was examined by immunoeytochemistry method and Western blotting. Results： （1） Immunocytochemistry detection targeting neuron specific enolase （NSE） and beta-Ⅲ tubulin confirmed that the cultured cells were neurons. （2）The expression of NF-κB P50 protein was significantly higher in OGD 4 h group than in control group（P〈0.05）; the expression continued to increase in OGD 4 h/R 2 h andOGD 4 h/R 6 h groups,and reached its peak 6 h after reoxygenation （P〈0. 01） ,then began to decrease,but the expression in OGD 4 h/R 12 h group was significantly different from that of the control group （P〈0.05） ; there was no significant difference between OGD 4 h/R 24 h group and control group. （3） The expression of NF-κB c-Rel protein was similar between OGD 4 h group and the control group; the expression increased in OGD 4 h/R 2 h and OGD 4 h/R 12 h groups and reached its peak 12 h after reoxygenation（P〈0.01）,and did not recover to the normal level in OGD 4 h/R 24 h group （P〈0.05）. Conclusion： Oxygen glucose deprivation/reoxygenation can activate NF-κB subunits P50 and c-Rel in the primary cortical neurons in rats in a time-associated manner.

关 键 词：NF-ΚB P50 C-REL 大脑皮质神经元 脑缺氧缺血 再灌注损伤 

分 类 号：R741.02[医药卫生—神经病学与精神病学]