MicroRNA let-7i regulates dendritic cells maturation targeting interleukin-10 via the Janus kinase 1-signal transducer and activator of transcription 3 signal pathway subsequently induces prolonged cardiac allograft survival in rats
MicroRNA let-7i regulates dendritic cells maturation targeting interleukin-10 via the Janus kinase 1-signal transducer and activator of transcription 3 signal pathway subsequently induces prolonged cardiac allograft survival in rats
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DOI：
10.1016/j.healun.2015.10.041
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发表时间：
2016-03-01
影响因子：
8.9
通讯作者：
Wu, Jian
中科院分区：
文献类型：
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作者：
Sun, Yong;Jin, Xiangyuan;Wu, Jian
背景：在本研究中，我们探讨了microRNA let-7i是否通过Janus kinase1信号转导和转录激活因子3(Janus kinase1-STAT3，JAK1-STAT3)信号通路调节靶向白介素10(IL-10)的树突状细胞成熟从而延长大鼠同种异体心脏移植物存活时间。方法：采用实时定量逆转录聚合酶链式反应、酶联免疫吸附试验和双荧光素酶试验验证IL-10是否是LET-7I作用的靶点，并用流式细胞仪和免疫组织化学方法检测调节性T细胞。Western印迹法检测JAK1、STAT3和磷酸化STAT3的表达。接受Lewis黑暗Agti心脏移植的受者输注磷酸盐缓冲液、脂多糖成熟的树突状细胞(MDCS)或let-7I抑制物MDCS。结果：与脂多糖-MDCS相比，LET-7I抑制剂-MDCS的IL-10信使RNA表达水平和IL-10的产生均显著增加。荧光素酶活性显示，IL-10荧光素酶报告基因的翻译水平被LET-7I模拟降低，而被LET-7I-抑制剂增加。MicroRNA let 7I抑制剂抑制DC成熟，但IL-10小干扰RNA可减弱这种抑制作用。MDC中JAK1、STAT3和磷酸化STAT3的表达可被LET-7I模拟抑制，而IL-10小干扰RNA的预处理可被LET-7I抑制剂上调。Lewis受体输注let-7i抑制剂-MDCS后，移植心脏存活时间显著延长。输注LET-7I抑制剂-MDCS的同种异体移植物细胞浸润较轻，移植物结构保存良好。抑制let-7i可增加Ichead box P3(+)调节性T细胞的CD4(+)CD25(+)，并调节体内和体外的细胞因子谱。结论：microRNA let-7i通过JAK1-STAT3途径调节DC的成熟和靶向IL-10的功能。此外，输注转LET-7I抑制剂的脂多糖诱导的骨髓间充质干细胞可延长同种异体心脏移植物S的存活时间并产生调节性T细胞。(C)2016年国际心肺移植学会。版权所有。
BACKGROUND: In this study, we investiglated whether microRNA let-7i regulates dendric cell maturation targeting interleukin-10 (IL-10) via the Janus kinase 1 signal transducer and activator of transcription 3 (JAK1-STAT3) signal pathway subsequently prolongs rat cardiac allograft survival.METHODS: Quantitative real-time reverse transcriptase polymerase chain reaction, enzyme linked immunosorbent assay, and dual-luciferase assay were performed to verify whether IL-10 was the target of let-7i, and regulatory T cells were assessed by flow cytometry and immunohistochemical study. Western blot was performed to detect JAK1, STAT3, and phosphorylated STAT3 expression. Lewis recipients of Dark Agouti hearts were transfused with phosphate-buffered saline, lipopolysaccharide (LPS)-mature dendritic cells (mDCs), or let-7i-inhibitor-mDCs. Allograft survival times were recorded, and histologic studies were performed.RESULTS: Expression of IL-10 messenger RNA level and production of IL-10 were increased in let-7i inhibitor-mDCs compared with LPS-mDCs. Luciferase activity showed that the translational level of the IL-10 luciferase reporter was decreased by let-7i mimic but increased by let-7i-inhibitor. MicroRNA let 7i inhibitor suppressed DC maturation; however, pretreatment of IL-10 small interfering RNA attenuated the suppression. Expression of JAK1, STAT3, and phosphorylated STAT3 in mDCs were suppressed by let-7i mimic, and pre-treatment of IL-10 small interfering RNA, however, were upregulated by let-7i inhibitor. Lewis recipients transfused with let-7i inhibitor-mDCs significantly prolonged Dark Agouti cardiac allograft survival. The allografts transfused with let-7i inhibitor-mDCs showed slight cell infiltration and significantly preserved graft structure. Inhibition of let-7i increased CD4(+)CD25(+)forIchead box P3(+) regulatory T cells and modulated cytokine profiles in vivo and in vitro.CONCLUSIONS: MicroRNA let-7i regulated DC maturation and function targeting IL-10 through the JAK1-STAT3 pathway. Moreover, transfusion of LPS-induced mDCs transfected with let-7i inhibitor induced prolonged cardiac allograft s'urvival and generated regulatory T cells. (C) 2016 International Society for Heart and Lung Transplantation. All rights reserved.