大鼠大脑皮质神经元氧糖剥夺/复氧后NF-κB P50/c-Rel对Bcl-xL表达的影响  

Influence of NF-κB P50/c-Rel on Bcl-x_L expression in primary cortical neurons of rats after oxygen glucose deprivation/reoxygenation

作　　者：何兰英[1] 罗勇[2] 

机构地区：[1]成都市第二人民医院神经内科,成都610017 [2]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》2011年第3期258-261,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：目的探讨脂质体(TransfastFastTM Transfection Reagent)介导转录因子NF-κB圈套物(decoy)寡聚脱氧核苷酸(oligodeoxynucleotide,ODNs)对Wistar大鼠大脑皮质神经元氧糖剥夺/复氧(OGD/R)后受NF-κB P50、c-Rel调控的抗凋亡因子Bcl-xL表达的影响。方法原代大脑皮质神经元体外培养,建立OGD/R模型,分为OGD 4 h/R 6 h组、NF-κB decoy ODNs组、无关decoy ODNs组、脂质体组;同时设立正常对照组。采用蛋白质印迹分析检测NF-κB P50、c-Rel蛋白表达;采用反转录-聚合酶链反应(RT-PCR)检测大鼠皮质神经元内Bcl-xL mRNA表达。结果蛋白质印迹分析显示OGD 4 h/R6 h组P50、c-Rel蛋白表达较正常组升高(P<0.01);转染NF-κB decoy ODNs后P50、c-Rel蛋白表达低于OGD 4 h/R 6 h组、脂质体组、无关decoy ODNs组(P<0.05);RT-PCR显示OGD 4 h/R 6 h组Bcl-xL mRNA表达较正常组升高(P<0.05);NF-κB decoy ODNs组Bcl-xL mRNA表达低于OGD 4 h/R 6 h组、脂质体组、无关decoy组(P<0.05)。结论 NF-κB decoy ODNs可有效抑制大脑皮质神经元OGD/R后受NF-κB亚基P50、c-Rel调控的抗凋亡因子Bcl-xL mRNA表达,这可能是NF-κB神经保护作用的部分机制。Objective To investigate the effect of TransfastFastTM Transfection Reagent-mediated transfection of NF-κB decoy oligodeoxynucleotides（ODNs） on expression of anti-apoptosis factor Bcl-xL in the primary cultured cortical neurons after oxygen glucose deprivation/reoxygenation（OGD/R）.Methods OGD/R model was established using primary cultured rat cortical neurons,and the model neurons were divided into 4 groups according to different treatments： OGD 4 h/R 6 h,NF-κB decoy ODNs,scrambled decoy ODNs and TransfastFastTM Transfection Reagent groups.Untreated neurons served as normal control.The protein expression of NF-κB P50-and c-Rel in the neurons was determined by Western blotting analysis;the expression of Bcl-xL mRNA was examined by RT-PCR.Results The expression of NF-κB P50 and c-Rel protein was significantly higher in the OGD 4 h/R 6 h than that in the normal control group（P〈0.01）;and the expression in NF-κB decoy ODNs group was significantly lower than that in the OGD 4 h/R 6 h,scrambled decoy ODNs and TransfastFastTM Transfection Reagent groups（P〈0.05）.RT-PCR results showed that Bcl-xL mRNA expression in OGD 4 h/R 6 h was significantly higher than that in the normal control group（P〈0.05）,and that in NF-κB decoy ODNs group was significantly lower than that in the OGD 4 h/R 6 h,scrambled decoy ODNs and TransfastFastTM Transfection Reagent groups（P〈0.05）.Conclusion NF-κB decoy ODNs can effectively inhibit NF-κB P50-and c-Rel-mediated Bcl-xL mRNA expression in primary cortical neurons,which may be one of the mechanisms for the neuroprotective function of NF-κB.

关 键 词：NF-ΚB 大脑皮质 神经元 氧糖剥夺/复氧 再灌注损伤 Bcl-xL 

分 类 号：R741.02[医药卫生—神经病学与精神病学]