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目的 :研究白介素 1(IL 1)受体相关激酶 2 (IRAK 2 )的反义寡核苷酸对IL 1生物学效应的影响。方法 :Lipofectin介导反义IRAK 2寡核苷酸转染人脐静脉内皮细胞。用SandwichELISA法检测核因子 kappaB(NF kappaB)的活化 ,竞争ELISA法测定PGI2 合成 ,细胞ELISA法检测细胞间粘附分子 1(ICAM 1)蛋白表达水平 ,以逆转录PCR法检测ICAM 1mRNA和I RAK 2mRNA表达水平。结果 :①反义IRAK 2寡核苷酸呈浓度 (1～ 4 μg·ml-1)和时间 (5～ 2 4h)依赖性地抑制NF kappaB活化、PGI2 合成和ICAM 1蛋白表达 ,反义IRAK 2寡核苷酸 3 μg·ml-1与细胞共孵育 8h时抑制效果最好。②反义IRAK 2寡核苷酸抑制ICAM 1mRNA表达。③反义IRAK 2寡核苷酸抑制IRAK 2mRNA表达。结论 :反义IRAK 2寡核苷酸通过抑制IRAK 2表达抑制IL 1生物活性 ,预示反义IRAK 2寡核苷酸是一种可供选择的抗炎新策略。
AIM: To investigate the effect of antisense oligonucleotide of interleukin 1 (IL-1) receptor-associated kinase 2 (IRAK 2) on the biological effects of IL-1. Methods: Lipofectin mediated antisense IRAK 2 oligonucleotide transfection of human umbilical vein endothelial cells. Sandwich ELISA was used to detect the activation of nuclear factor kappaB (NF kappaB), PGI2 was detected by competition ELISA, the expression of ICAM 1 was detected by ELISA, ICAM 1 mRNA and I RAK were detected by reverse transcription polymerase chain reaction 2 mRNA expression level. RESULTS: Antisense IRAK 2 oligodeoxynucleotides inhibited NF-kappaB activation, PGI2 synthesis and ICAM-1 protein expression in a concentration-dependent manner (1-4 h · min-1) and time- (5-24 h) 2 3μg · ml-1 oligonucleotide incubated with cells 8h best inhibitory effect. ② Antisense IRAK 2 oligonucleotide inhibited ICAM 1 mRNA expression. ③ antisense IRAK 2 oligonucleotide inhibited IRAK 2 mRNA expression. Conclusion: The antisense IRAK 2 oligonucleotide inhibits the biological activity of IL 1 by inhibiting the expression of IRAK 2, indicating that antisense IRAK 2 oligonucleotide is an alternative anti-inflammatory strategy.