IL-1受体相关激酶-1和-2协同调控IL-1诱导的AP-1活化

IL-1 receptor associated kinase-1 and-2 synergistically activate AP-1

目的研究白介素 - 1受体相关激酶 - 1(IRAK- 1)和 IRAK- 2在白介素 - 1(IL - 1)诱导 AP- 1活化中的作用。方法L ipofectin介导反义 IRAK- 1寡核苷酸和反义 IRAK- 2寡核苷酸转染 Hep G2细胞。用逆转录 PCR法检测 IRAK - 1和 IRAK- 2m RNA表达水平 ;Western blot分析 IRAK- 1和 IRAK - 2蛋白表达水平。以 Sandwich EL ISA法检测 AP- 1的活化。结果反义IRAK- 1寡核苷酸和反义 IRAK- 2寡核苷酸通过抑制各自靶基因 m RNA和蛋白表达抑制 IL- 1诱导的 AP- 1活化 ;反义 IRAK-1寡核苷酸与反义 IRAK- 2寡核苷酸共转染 Hep G2细胞对 AP- 1的抑制作用较两者单独转染明显增强。结论 IRAK- 1和 I-RAK- 2在调控白介素 - 1诱导的 AP- 1活化时协同作用。

ObjectiveTo investigate whether interleukin 1 receptor associated kinase 1 (IRAK 1) and IRAK 2 are functionally combined or redundant but differentially expressed in interleukin 1(IL 1) induced activator protein 1(AP 1) activation. Methods Phosphorothioate oligonucleotides(ODN) were designed antisense to sequences of IRAK 1 or IRAK 2. Antisense IRAK 1 ODN or antisense IRAK 2 ODN was delivered by lipofectin encapsulation into cultured HepG2 cells. IRAK 1 and IRAK 2 mRNA expression were assayed by semiquantitative reverse transcription PCR. IRAK 1 and IRAK 2 protein expression were detected by western blot. The levels of activator protein 1(AP 1) were measured by sandwich ELISA. Results Antisense IRAK 1 ODN and antisense IRAK 2 ODN blocked IRAK 1 and IRAK 2 expression, respectively. As a result, antisense IRAK 1 ODN or antisense IRAK 2 ODN inhibited IL 1 induced AP 1 activation. Cotransfection of antisense IRAK 1 ODN 4 μg with antisense IRAK 2 ODN 4 μg resulted in an additive inhibition of AP 1 activation. Conclusion IRAK 1 regulates IL 1 stimulated AP 1 activation in cooperation with IRAK 2. [