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为研究IL 1R相关激酶 1(IRAK 1)和磷脂酰肌醇 3 激酶 (PI 3 激酶 )在IL 1诱导NF κB活化中的作用 ,本文采用Lipo fectin介导反义IRAK 1寡核苷酸或反义PI 3 激酶寡核苷酸转染HEpG2细胞后 ,用Western杂交检测IRAK 1和PI 3 激酶表达水平 ,以SandwichELISA法检测NF κB的活化。结果表明 ,①反义IRAK 1寡核苷酸和反义PI3 激酶寡核苷酸分别抑制IRAK 1和PI3 激酶表达 ;②反义IRAK 1寡核苷酸或反义PI 3 激酶寡核苷酸部分抑制NF κB活化 ;③与反义IRAK 1寡核苷酸或反义PI 3 激酶寡核苷酸单独转染HEpG2细胞相比 ,反义IRAK 1寡核苷酸和反义PI 3 激酶寡核苷酸共转染HEpG2细胞对NF κB的抑制作用明显增强。表明IRAK 1或PI3 激酶调控NF κB活化但不能完全激活NF κB ,IRAK 1和PI3 激酶在调控NF κB活化时协同作用。
In order to investigate the role of IL-1R-related kinase 1 (IRAK 1) and phosphatidylinositol 3-kinase (PI 3-kinase) in IL-1-induced activation of NF-κB, Lipofectin was used to mediate antisense IRAK 1 oligonucleotide or anti- After HEpG2 cells were transfected with sense PI 3 kinase oligonucleotides, the expression of IRAK 1 and PI 3 kinase was detected by Western blotting. The activation of NF κB was detected by Sandwich ELISA. The results showed that: (1) the antisense IRAK 1 oligonucleotide and the antisense PI3 kinase oligonucleotide inhibited the expression of IRAK 1 and PI3 kinase respectively; (2) the antisense IRAK 1 oligonucleotide or antisense PI 3 kinase oligonucleotide Inhibition of NF κB activation; ③ antisense IRAK 1 oligonucleotide or antisense PI 3 kinase oligonucleotide transfected HEpG2 cells alone compared to antisense IRAK 1 oligonucleotide and antisense PI 3 kinase oligonucleotide The inhibitory effect of acid co-transfected HEpG2 cells on NF κB was significantly enhanced. It is suggested that IRAK 1 or PI3 kinase regulates NF-κB activation but not NF-κB. IRAK 1 and PI3-kinase cooperate synergistically in the regulation of NF-κB activation.