Adenosine promotes Foxp3 expression in Treg cells in sepsis model by activating JNK/AP-1 pathway
Adenosine promotes Foxp3 expression in Treg cells in sepsis model by activating JNK/AP-1 pathway
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腺苷通过激活JNK/AP-1通路促进脓毒症模型Treg细胞中Foxp3的表达
DOI：
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Tao Yang
中科院分区：
文献类型：
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作者：
Rui Bao;Jiong Hou;Yan Li;Jinjun Bian;Xiaoming Deng;Xiaoyan Zhu;Tao Yang
目的：叉头/翼螺旋转录因子p3 （Foxp3）在败血症期间CD4+CD25+Treg细胞中升高；然而，相关机制尚不清楚。我们的研究旨在探讨脓毒症期间Treg细胞中Foxp3高表达的可能分子机制。方法：采用盲肠结扎穿刺法（CLP）诱导脓毒症。从外周血中分离CD4+CD25+Treg细胞，流式细胞术（FCM）鉴定。分别用或不加腺苷、腺苷激动剂、腺苷拮抗剂、SMAD家族成员3 （Smad3）激动剂（转化生长因子(TGF)-β1）或C-Jun n -末端激酶（JNK）抑制剂培养Treg细胞。检测Foxp3和激活蛋白1 （AP-1）的表达水平。然后通过染色质免疫沉淀（ChIP）法评估c-Fos或c-Jun与Foxp3启动子的结合，并通过定量实时PCR （qRT-PCR）进行定量。用siRNA转染c-Fos、Fra-2、c-Jun或JunD后，检测Foxp3的mRNA和蛋白水平。结果：腺苷和JNK的药理抑制均可降低Foxp3蛋白水平。JNK/AP-1的激活与CD4+CD25+Treg细胞中Foxp3蛋白水平升高有关。AP-1调节Treg细胞中Foxp3启动子的活性，而c-Fos或c-Jun活性的诱导导致Foxp3基因转录的升高。c-Fos、Fra-2、c-Jun或JunD水平的下调也会降低Foxp3的表达。结论：腺苷在Foxp3高表达中起重要作用。腺苷通过JNK/AP-1途径促进脓毒症期间Treg细胞Foxp3的表达。
Objectives: Forkhead/winged helix transcription factor p3 (Foxp3) increases in CD4+CD25+Treg cells during sepsis; however, related mechanisms are unclear. Our study aimed to explore the possible molecular mechanisms of high expression of Foxp3 in Treg cells during sepsis. Methods: Sepsis was induced by cecal ligation and puncture (CLP) method. CD4+CD25+Treg cells were isolated from peripheral blood and identified by flow cytometry (FCM). Treg cells were cultured with or without adenosine, adenosine agonist, adenosine antagonist, SMAD family member 3 (Smad3) agonist (transforming growth factor (TGF)-β1), or C-Jun N-Terminal Kinase (JNK) inhibitor.Expression levels of Foxp3 and activator protein 1 (AP-1) were determined. The binding of c-Fos or c-Jun to the Foxp3 promoter was then evaluated by the chromatin immunoprecipitation (ChIP) assay and quantified by quantitative real-time PCR (qRT-PCR). The mRNA and protein levels of Foxp3 were determined after transfection with siRNA against c-Fos, Fra-2, c-Jun or JunD. Results: Pharmacological inhibition of both adenosine and JNK reduced Foxp3 protein levels. JNK/AP-1 activation was involved in increased levels of Foxp3 protein in CD4+CD25+Treg cells. AP-1 regulated activity of Foxp3 promoter in Treg cells, and the induction of c-Fos or c-Jun activity leads to elevated transcription of Foxp3 gene. Knockdown of c-Fos, Fra-2, c-Jun, or JunD levels also reduced Foxp3 expression. Conclusion: We confirm that adenosine plays significant roles in the high expression of Foxp3. Adenosine promotes Foxp3 expression in Treg cells during sepsis via JNK/AP-1 pathway.