| 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 4 h组、OGD 4 h/R 2 h组、OGD 4 h/R 6 h组、OGD 4 h/R 12 h组、OGD 4 h/R 24 h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 4 h组NF-κB P50蛋白表达开始增加明显高于正常组(P<0.05),OGD 4 h/R 2 h组、OGD 4 h/R 6 h组NF-κB P50蛋白表达持续性升高,并在复氧后6 h达高峰(P<0.01),此后逐渐下降,至OGD 4 h/R 12 h时与正常组有显著差异(P<0.05),OGD 4 h/R 24 h 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 4 h组与正常组无差异(P>0.05),OGD 4 h/R 2 h至OGD 4 h/R 12 h表达持续增加,并在复氧后 12 h达高峰(P<0.01),OGD 4 h/R 24 h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。 |
| 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 |
| DOI：10.3724/SP.J.1008.2008.01333 |
| 投稿时间：2008-04-27修订日期：2008-09-26 |
| 基金项目:重庆市卫生局资助项目(渝卫科教\[2003\]61号,03-2-179). |
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| Expression of NF-κB subunits P50 and c-Rel protein in primary cortical neurons after oxygen glucose deprivation/reoxygenation |
| HE Lan-ying,LUO Yong* |
| (Department of Neurology,Chongqing Key Laboratory of Neurology,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China) |
| Abstract: |
| 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 immunocytochemistry 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. |
| Key words: NF-κB P50,c-Rel cortical neurons brain hypoxia-tschemia reperfusion injury |
