| 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/R 6 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。 |
| 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 |
| DOI：10.3724/SP.J.1008.2011.0258 |
| 投稿时间：2010-12-17修订日期：2011-01-12 |
| 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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| Influence of NF-κB P50/c-Rel on Bcl-xL expression in primary cortical neurons of rats after oxygen glucose deprivation/reoxygenation |
| HE Lan-ying1, LUO Yong2* |
| (1. Department of Neurology, the Second People’s Hospital of Chengdu, Chengdu 610017, Sichuan, China; 2. Department of Neurology,the First Affiliated Hospital, Chongqing Medical University, Chongqing Key Laboratory of Neurology, Chongqing 400016, China) |
| Abstract: |
| ObjectiveTo 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). MethodsOGD/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. ResultsThe 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). ConclusionNF-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. |
| Key words: NF-kappaB cerebral cortex neurons oxygen glucose deprivation/reoxygenation |
