脂多糖刺激干扰素调节因子3促进小鼠腹腔巨噬细胞白介素-17表达

Interferon regulatory factor 3 promotes the expression of interleukin-17 in mouse peritoneal macrophages stimulated by lipopolysaccharide

目的:探讨干扰素调节因子3(IRF3)对脂多糖(LPS)刺激小鼠腹腔巨噬细胞促进白细胞介素-17(IL-17)表达的初步作用机制。方法:小鼠腹腔注射3%巯基乙酸肉汤,3 d后提取C57BL/6野生和IRF3基因敲除小鼠的腹腔巨噬细胞,培养过夜后添加LPS。收集细胞培养上清液,酶联免疫吸附试验检测细胞因子IL-17和白细胞介素-6(IL-6)的表达;提取细胞蛋白质,免疫印迹检测细胞核因子κB抑制蛋白(IκB)α、IRF3、磷酸化信号转导子和转录激活子3(STAT3)的蛋白水平。结果:LPS刺激小鼠腹腔巨噬细胞后,IRF3可显著地促进IL-17的产生,同时伴随着IL-6的产生、IκBα蛋白的降解及STAT3的磷酸化。结论:IRF3可能通过促进IL-6的产生,间接地激活STAT3的磷酸化,进而促进IL-17的产生。

Objective:To explore the effect of interferon regulatory factor 3(IRF3)on the expression of interleukin-17(IL-17)in mouse peritoneal macrophages stimulated by lipopolysaccharide(LPS).Methods:Mice were intraperitoneally injected with 3%thioglycolic acid broth.Three days later,the peritoneal macrophages of C57BL/6 wild and IRF3 gene knockout mice were extracted and cultured overnight,then stimulated with LPS.Enzyme linked immunosorbent assay was used to detect the expression of cytokines IL-17 and interleukin-6(IL-6)in cell culture supernatant.Western blotting was used to detect the expression of inhibitor of nuclear factor kappa B(IκB)α,IRF3,phosphorylated signal transducer and transcription activator 3(STAT 3)in cells.Results:After LPS stimulation,the expression of IL-17 in mouse peritoneal macrophages was significantly promoted by IRF3,accompanied by the production of IL-6,the degradation of IκBα,and STAT3 phosphorylation.Conclusions:IRF3 may indirectly activate STAT3 phosphorylation by promoting the production of IL-6,thereby promoting the production of IL-17.