摘要
目的研究白介素 1(IL 1)受体相关激酸 2(IRAK 2)在IL 1诱导NF κB活化中的作用。方法分别以逆转录PCR法和Western杂交法 ,检测lipofectin介导的反义I RAK 2寡核苷酸转染的人脐静脉内皮细胞中 ,IRAK 2mR NA和蛋白表达水平。用夹心ELISA法检测NF κB的活化。结果反义IRAK 2寡核苷酸可阻断IRAK 2mRNA的翻译 ,从而抑制IRAK 2蛋白的表达 ,抑制率达45% ;反义IRAK 2寡核苷酸抑制IL 1诱导的NF κB活化 ,具有剂量和时间依赖性 ,3μg与8h时的抑制效果最好。结论IRAK 2可调控IL 1诱导的NF κB活化。
Aim To investigate the role of interleukin 1(IL 1) receptor associated kinase 2(IRAK 2) in IL 1 induced nuclear factor κB(NF κB) activation. Methods Antisense phosphorothioate oligonucleotide(ODN) was designed to sequences of IRAK 2. Antisense IRAK 2 ODN was delivered by lipofectin encapsulation into cultured endothelial cells. IRAK 2 mRNA and protein expression were detected by semiquantitative reverse transcription PCR and Western blot, respectively. The level of NF κB was measured by sandwich ELISA. Results Antisense IRAK 2 ODN blocked IRAK 2 mRNA translation and protein expression was inhibited. As a result, antisense IRAK 2 ODN inhibited IL 1 induced NF κB activation in a dose(1 4 μg) and time(5 24 h) dependent manner. When the cells were treated with 3 μg of antisense IRAK 2 ODN for 8 h, a maximum inhibition was achieved. Conclusion IRAK 2 might be necessary for IL 1 stimulated NF κB activation.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
2000年第2期128-130,共3页
Chinese Journal of Cellular and Molecular Immunology
关键词
反义寡核苷酸
核因子-KB
interleukin 1
interleukin 1 receptor associated kinase 2
antisense oligonucleotide
nuclear factor κB
