Swine IRF3/IRF7 attenuates inflammatory responses through TLR4 signaling pathway.
Swine IRF3/IRF7 attenuates inflammatory responses through TLR4 signaling pathway.
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DOI：
10.18632/oncotarget.18740
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发表时间：
2017-09-22
期刊：
影响因子：
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通讯作者：
Li HP
中科院分区：
文献类型：
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作者：
Chen PG;Guan YJ;Zha GM;Jiao XQ;Zhu HS;Zhang CY;Wang YY;Li HP
为了探讨IRF 3/IRF 7在炎症反应中的作用，我们研究了猪IRF 3/IRF 7对猪肾上皮细胞株PK 15中TLR 4信号通路和炎症因子表达的影响。我们成功构建了真核表达载体PB-IRF 3和PB-IRF 7，并将其转染PK 15细胞，在荧光显微镜下观察到GFP的存在。此外，还采用RT-PCR检测转染效率。我们发现IRF 3/IRF 7在PK 15细胞中高效过表达。此外，我们还通过RT-PCR检测了IRF 3/IRF 7对TLR 4信号通路和炎症因子的影响。用脂多糖（LPS）单独或与TBK 1抑制剂（LiCl）组合处理转染的细胞。我们发现，IRF 3/IRF 7增强IFNα的产生，并降低IL-6 mRNA的表达。阻断TBK 1通路可抑制IFNα mRNA的变化，但对IL-6 mRNA的变化无影响。这说明IRF 3/IRF 7通过TLR 4/TBK 1信号通路促进IFNα的产生并发挥抗炎作用，而IRF 3/IRF 7不依赖于TBK 1通路而降低IL-6的表达。MyD 88、TRAF 6、TBK 1和NFκB mRNA变化趋势在所有处理中相似。LPS可使PBR 3/PBR 7和PBv细胞MyD 88、TRAF 6、TBK 1和NFκB mRNA表达增加，而LiCl可阻断LPS的作用。PBR 3/PBR 7细胞中上述四种因子的水平均高于PBv细胞。这些结果表明IRF 3/IRF 7通过TLR 4信号通路调节炎症反应。在PK 15细胞中过表达猪IRF 3/IRF 7可诱导I型干扰素的产生，并通过TLR 4信号通路减轻炎症反应。
To explore the role of IRF3/IRF7 during inflammatory responses, we investigated the effects of swine IRF3/IRF7 on TLR4 signaling pathway and inflammatory factors expression in porcine kidney epithelial PK15 cell lines. We successfully constructed eukaryotic vectors PB-IRF3 and PB-IRF7, transfected these vectors into PK15 cells and observed GFP under a fluorescence microscope. In addition, RT-PCR was also used to detect transfection efficiency. We found that IRF3/IRF7 was efficiently overexpressed in PK15 cells. Moreover, we evaluated the effects of IRF3/IRF7 on the TLR4 signaling pathway and inflammatory factors by RT-PCR. Transfected cells were treated with lipopolysaccharide (LPS) alone, or in combination with a TBK1 inhibitor (LiCl). We revealed that IRF3/IRF7 enhanced IFNα production, and decreased IL-6 mRNA expression. Blocking the TBK1 pathway, inhibited the changes in IFNα, but not IL-6 mRNA. This illustrated that IRF3/IRF7 enhanced IFNα production through TLR4/TBK1 signaling pathway and played an anti-inflammatory role, while IRF3/IRF7 decreased IL-6 expression independent of the TBK1 pathway. Trends in MyD88, TRAF6, TBK1 and NFκB mRNA variation were similar in all treatments. LPS increased MyD88, TRAF6, TBK1 and NFκB mRNA abundance in PBR3/PBR7 and PBv cells, while LiCl blocked the LPS-mediated effects. The levels of these four factors in PBR3/PBR7 cells were higher than those in PBv. These results demonstrated that IRF3/IRF7 regulated the inflammatory response through the TLR4 signaling pathway. Overexpression of swine IRF3/IRF7 in PK15 cells induced type I interferons production, and attenuated inflammatory responses through TLR4 signaling pathway.