脂多糖(LPS)对小鼠Raw 264.7细胞Irak1bp1表达的影响

Effects of lipopolysaccharide (LPS) on the expression of Irak1bp1 in mouse Raw 264.7 cells

目的研究脂多糖(lipopolysaccharide,LPS)刺激小鼠Raw264.7细胞IL-1受体相关激酶1结合蛋白1(IL-1 receptor associated kinase 1 binding protein1,Irak1bp1)的表达情况。方法体外培养的小鼠Raw264.7细胞随机分为2组:A组为正常对照组,B组为LPS刺激组。LPS刺激小鼠Raw264.7细胞6h后收集细胞及上清液。采用实时荧光定量PCR法检测IL-6及Irak1bp1mRNA水平;采用ELISA分析培养上清液中IL-6含量;采用Western blot方法检测Irak1bp1蛋白质水平。结果LPS刺激组炎症因子IL-6mRNA及蛋白质水平较对照组明显增高(P<0.01),Irak1bp1mRNA及总蛋白质水平高于对照组(P<0.05);胞质内Irak1bp1蛋白质水平与对照组相比无明显变化,而胞核内Irak1bp1蛋白质水平较正常对照组增加(P<0.01)。结论LPS能够诱导Raw264.7细胞核内Irak1bp1表达,且该分子的表达上调可能与细胞的炎性因子释放相关。

Objective To investigate the expression of IL-1 receptor associated kinase 1 binding protein 1 (Irak1bp1) in lipopolysaccharide (LPS)-stimulated mouse Raw 264.7 cells. Methods Cultured mouse Raw 264.7 cells were devided randomly into two groups:normal control (A) group and LPS stimulation (B) group.Cells and supernatants were collected after mouse Raw 264.7 cells treated with LPS for 6 h.The expressions of IL-6 and Irak1bp1 mRNA were detected by real-time PCR.The secretion of IL-6 in cell supernatants was analyzed by ELISA.The expression of Irak1bp1 protein was detected by Western blot. Results The expression and secretion of IL-6 mRNA and protein were significantly higher than the control group ( P <0.01).The expression of Irak1bp1 mRNA and total Irak1bp1 protein was significantly higher than the control group ( P <0.05).The expression of Irak1bp1 protein in cytoplasm was not significantly changed compared with normal,while in cell nucleus it significantly increased ( P <0.01). Conclusions LPS stimulation can induce the expression of nuclear Irak1bp1 protein,which may be associated with inflammatory releases in Raw 264.7 cells.