利用siRNA抑制NF-κB p65蛋白表达对EOMA细胞中TF表达影响  

The inhibiting effect of siRNA targeting NF - kB p65 to the expression of TF in EOMA cell

作　　者：周晓彤[1] 沈振亚[2] 余云生[2] 叶文学[2] 黄浩岳[2] 滕小梅[2] 

机构地区：[1]无锡市人民医院心血管外科,江苏无锡214023 [2]苏州大学心血管病研究所,江苏苏州215006

出　　处：《徐州医学院学报》2012年第11期701-704,共4页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金九五重大课题（39993430）

摘　　要：目的探讨针对小鼠EOMA细胞核冈子KB（NF—KB）的小干扰RNA（siRNA）对细胞c}t组织因子（TF）表达的抑制作用。方法设计并化学合成针对小鼠NF—KBp65的siRNA，按照实验分组分别转染小鼠EOMA细胞，转染24h后，在相应细胞中加入肿瘤坏死因子α（TNF-α，终浓度为20mg／L），于此后6h分别提取各组细胞总蛋白及RNA，检测细胞内NF—KBp65蛋白表达和TF的mRNA水平。结果TNF-α．刺激6h后，与空白对照组相比，阴性对照组、scramblesiRNA组细胞内NF—KBp65蛋白表达和TFmRNA水半增加，而siRNA组则明娃受到抑制；TNF组、TNF＋scramblesiRNA组细胞内NF—KBp65蛋门表达和TF mRNA水平明显增加，TNF＋siRNA组细胞内NF—KBp65蛋白表达和TFmRNA水平较卒白对照组有所增加，但明显低下TNF纽、TNF＋scramblesiRNA组（P〈0．01）。结论利用siRNA抑制NF—KBp65蛋白表达可以有效地抑制NF—KB下游靶慕因TF的表达。Objective To explore the inhibiting effect of siRNA targeting NF - KB p65 to the expression of tissue factor （TF） in EOMA cell. Methods EOMA cell was devided in to seven groups, siRNA directed against mouse NF - KB p65 was designed and was chemically synthesized. The treatment of TNF -α was performed 24 h after transleetion, and 6 h later, the expression level of NF - KB p65 protein was evaluated by Western blot, the expression level of TF mR- NA was evaluated by RT - PCR in each groups. Results The level of NF - KB p65 protein and TF in negative control group and scramble siRNA group was increased at 6 h, but was significantly suppressed in siRNA group. Due to the stim- ulation of TNF - α, the expression of NF - KB p65 protein and TF were remarkably enhanced in TNF group and TNF ＋ scramble siRNA group. In TNF ＋ siRNA group, the levels of NF - KB 1365 protein and mRNA of TF was higher than that in blank control group, but were lower than that in TNF group and TNF ＋ scramble siRNA group （P 〈0.01 ）. Conclution The inhibition of activation of NF - KB p65 by siRNA could efficiently suppress the TNF - α induced expression of TF in EOMA cell.

关 键 词：小干扰RNA 血管内皮细胞 核因子-KB 组织因子 延迟性排斥反应 

分 类 号：R392.4[医药卫生—免疫学]