MiR-21 Regulates TNF-α-Induced CD40 Expression via the SIRT1-NF-κB Pathway in Renal Inner Medullary Collecting Duct Cells
MiR-21 Regulates TNF-α-Induced CD40 Expression via the SIRT1-NF-κB Pathway in Renal Inner Medullary Collecting Duct Cells
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DOI：
10.1159/000455981
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Tian, Zhenjun
中科院分区：
文献类型：
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作者：
Lin, Qinqin;Geng, Yuanwen;Tian, Zhenjun
背景/目的：最近的研究表明microRNA-21（miR-21）参与了肾脏疾病相关的炎症反应。Sirtuin 1（SIRT 1）通过对抗炎症发挥肾脏保护作用。CD 40的激活触发炎症，参与肾脏炎症和损伤。然而，miR-21、SIRT 1和CD 40之间的关系仍然难以捉摸。研究方法：采用免疫组化、小干扰RNA（siRNA）转染、实时荧光定量PCR和蛋白质印迹等方法对原代培养的肾内髓集合管（IMCD）细胞进行形态学、功能学和分子机制的研究。结果：TNF-α诱导IMCD细胞中miR-21、CD 40和乙酰化NF-κ Bp 65（Ac-p65）的表达，并降低SIRT 1的表达。在TNF-α诱导的IMCD细胞中，miR-21模拟物增加了SIRT 1的表达，减弱了Ac-p65和CD 40的表达，并且在miR-21抑制剂的作用下观察到了相应的变化。SIRT 1过表达或SRT 1720激活可降低TNF-α诱导的CD 40和Ac-p65表达，SIRT 1 siRNA或抑制剂Ex 527和sirtinol可逆转该作用，NF-κ B siRNA预处理可增强该作用。进一步研究发现，miR-21对Ac-p6和CD 40表达的抑制作用可被SIRT 1 siRNA预处理所阻断。结论：目前的研究表明，miR-21通过SIRT 1-NF-κ B信号通路抑制TNF-α诱导的IMCD细胞中CD 40的表达，这为理解miR-21的抗炎作用提供了新的见解。(C)2017作者（s）由S. Karger AG，巴塞尔
Background/Aims: Recent studies have indicated that microRNA-21 (miR-21) is involved in the inflammatory response in relation to renal disease. Sirtuin1 (SIRT1) exerts renoprotective properties by counteracting inflammation. The activation of CD40 triggers inflammation that participates in renal inflammation and injury. The relationship between miR-21, SIRT1 and CD40, however, remains elusive. Methods: Immunohistochemistry, small-interfering RNA (siRNA) transfection, quantitative real-time PCR and western blotting were applied to assess the morphological, functional and molecular mechanisms in primary cultured renal inner medullary collecting duct (IMCD) cells. Results: TNF-alpha induced miR-21, CD40 and acetylated-NF-kappa Bp65 (Ac-p65) expressions and reduced SIRT1 expression in IMCD cells. miR-21 mimics increased SIRT1 expression and attenuated Ac-p65 and CD40 expressions in TNF-alpha-induced IMCD cells, and the corresponding changes were observed with a miR-21 inhibitor. SIRT1 overexpression or activation by SRT1720 diminished TNF-alpha-induced CD40 and Ac-p65 expressions, which was reversed by SIRT1 siRNA or the inhibitors Ex527 and sirtinol and augmented by pretreatment with NF-kappa B siRNA. Further study found that the inhibitory effect of miR-21 on Ac-p6 and CD40 expressions was impeded by pretreatment with SIRT1 siRNA. Conclusion: The present study indicates that miR-21 inhibits TNF-alpha-induced CD40 expression in IMCD cells via the SIRT1-NF-kappa B signalling pathway, which provides new insight in understanding the anti-inflammatory effect of miR-21. (C) 2017 The Author(s) Published by S. Karger AG, Basel