IL-1 预刺激对反义IRAK-1 寡核苷酸抑制NF-κB活化的影响

The Effects of IL-1-pretreatment on Inhibition of IL-1-induced NF-_κB Activation by Antisense IRAK-1O ligonucleotide

实验分为白介素－１（ＩＬ－１）预处理组和非预处理组，脂质体介导反义ＩＬ－１ 受体相关激酶－１（ＩＲＡＫ－１）寡核苷酸（ＯＤＮ） 转染ＨｅｐＧ２ 细胞，用ｗ ｅｓｔｅｒｎ 杂交分析ＩＲＡＫ－１ 表达水平，以夹心酶联免疫吸附测定法检测ＮＦ－κＢ含量． 结果表明，ＩＬ－１ 非预处理组反义ＩＲＡＫ－１ ＯＤＮ不能抑制ＩＲＡＫ－１ 表达和ＮＦ－κＢ活化，而预处理组ＩＲＡＫ－１ 表达和ＮＦ－κＢ活化受到明显抑制． 反义ＩＲＡＫ－１ ＯＤＮ 对ＮＦ－κＢ活化的抑制作用具有时间（５～２４ ｈ）和剂量（１～８ μｇ）依赖性． 说明ＩＬ－１ 预刺激在反义ＩＲＡＫ－１ ＯＤＮ抑制ＩＬ－１ 诱导的ＮＦ－κＢ活化中起决定性作用．

Antisense interleukin 1 (IL 1) receptor associated kinase 1(IRAK 1) oligonucleotide (ODN) was delivered by lipofectin encapsulation into HepG2 cells. The levels of NF κB were assayed by sandwich enzyme linked immunosorbent assay. The results showed that antisense IRAK 1 ODN did not inhibit IRAK 1 expression and IL 1 induced NF κB activation without IL 1 pretreatment. However, IRAK 1 expression and NF κB activation were significantly inhibited by antisense IRAK 1 ODN if the cells were pretreated with IL 1. Antisense IRAK 1 ODN inhibited NF κB activation in a time (5～24 h) and dose (1～8 μg) dependent manner. Therefore, IL 1 pretreatment is necessary for inhibi tion of IL 1 stimulated NF κB activation by antisense IRAK 1 ODN.