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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。
Objective To investigate the role of interleukin - 1 receptor - associated kinase - 1 (IRAK - 1) and IRAK - 2 in the activation of AP-1 induced by interleukin - 1 (IL - 1). Methods L ipofectin mediated transfection of Hep G2 cells with antisense IRAK-1 and antisense IRAK-2 oligonucleotides. The expression of IRAK - 1 and IRAK - 2m RNA was detected by RT - PCR. The protein expression of IRAK - 1 and IRAK - 2 was analyzed by Western blot. AP-1 activation was assayed by Sandwich EL ISA. Results The antisense IRAK-1 oligonucleotide and the antisense IRAK-2 oligonucleotide inhibited IL-1-induced AP-1 activation by inhibiting the respective target gene mRNA and protein expression; antisense IRAK-1 oligonucleotide Inhibition of AP-1 by Hep G2 cells co-transfected with antisense IRAK-2 oligonucleotide was significantly enhanced compared with that of Hep G2 cells alone. Conclusions IRAK-1 and I-RAK-2 are synergistic in the regulation of interleukin-1-induced activation of AP-1.