论文部分内容阅读
目的设计合成靶向哑铃形寡核苷酸(oligodeoxynucleotides,ODNs)圈套,检测其对大鼠大脑皮质神经元氧糖剥夺/复氧(OGD/R)后TNF-α表达的抑制效应。方法原代大脑皮质神经元体外培养,建立氧糖剥夺/复氧模型,以正常组、OGD4h/R6h组、NF-κB decoy ODNs组、无关decoy ODNs组、脂质体组为实验组,采用Western blot检测NF-κB p65蛋白表达,分别采用免疫细胞化学和逆转录-聚合酶链反应法(RT-PCR)检测TNF-α蛋白和mRNA表达。结果Westernblot检测显示NF-κB decoy ODNs组NF-κB P65蛋白表达明显低于OGD4h/R6h组、脂质体组、无关decoy ODNs组(P<0.05),与OGD4h/R6h组、脂质体组、无关decoy组相比NF-κB decoy ODNs组TNF-α蛋白和mRNA表达明显降低(P<0.05);而脂质体组、无关decoy ODNs组对NF-κB P65蛋白、TNF-α蛋白和mRNA表达未见抑制(P>0.05)。结论靶向性NF-κB的哑铃形圈套ODNs可有效抑制大脑皮质神经元OGD/R后炎症因子TNF-αmRNA及蛋白的表达。
OBJECTIVE: To design and synthesize a snakehead-targeting oligodeoxynucleotides (ODNs) scaffold to detect the inhibitory effect of TNF-α after OGD / R on rat cerebral cortical neurons. Methods Primary cultured rat cerebral cortical neurons were cultured in vitro. OGD4h / R6h group, NF-κB decoy ODNs group, decoy ODNs unrelated group and liposome group were used as experimental groups. The expression of NF-κB p65 protein was detected by Western blot. The expressions of TNF-α protein and mRNA were detected by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Results Western blot showed that the expression of NF-κB P65 in NF-κB decoy ODNs group was significantly lower than that in OGD4h / R6h group, liposome group and decoy ODNs group (P <0.05). Compared with OGD4h / R6h group, The expression of TNF-αprotein and mRNA in the decoy ODNs group was significantly lower than that in the decoy ODNs group (P <0.05). However, the expression of NF-κB P65 protein, TNF-αprotein and mRNA in the decoy ODNs group No inhibition (P> 0.05). Conclusion Dumbbell-shaped ODNs targeting NF-κB can effectively inhibit the expression of TNF-α mRNA and protein in inflammatory cells after OGD / R in cerebral cortical neurons.