反义IRAK-2寡核苷酸抑制白介素-1生物学效应

Inhibition of antisense IRAK-2 oligodeoxynucleotide on the biological effects of interleukin-1

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。

OBJECTIVE: To study the effects of antisense interleukin 1 (IL 1) receptor associated kinase 2 (IRAK 2) oligodeoxynucleotide (ODN) on the cellular response to IL 1 METHODS: Antisense IRAK 2 ODN was delivered by lipofectin encapsulation into cultured endothelial cells The levels of nuclear factor kappaB (NF kappaB), PGI 2 release and surface expression of intracellular adhesion molecule 1 (ICAM 1) were measured by sandwich ELISA, competitive ELISA, ELISA on cells in situ, respectively ICAM 1 and IRAK 2 mRNAs were detected by semiquantitative reverse transcription PCR RESULTS: ① Antisense IRAK 2 ODN inhibited IL 1 induced NF kappaB activation, PGI 2 release and surface expression of ICAM 1 in a concentration (1～4 μg·ml -1 ) and time (5～24 h) dependent fashion A maximum inhibition of NF kappaB activation, PGI 2 release or surface expression of ICAM 1 occurred when the cells were incubated with 3 μg·ml -1 of antisense IRAK 2 ODN for 8 h ② Antisense IRAK 2 ODN inhibited IL 1 induced ICAM 1 mRNA expression ③ Antisense IRAK 2 ODN inhibited IRAK 2 mRNA expression CONCLUSION: These data demonstrated that antisense IRAK 2 ODN inhibited biological effects of IL 1 via inhibition of IRAK 2 expression, suggesting that antisense IRAK 2 ODN may share a role in the design of antiinflammatory therapeutics