Diazoxide potentiates mesenchymal stem cell survival via NF-κB-dependent miR-146a expression by targeting Fas
Diazoxide potentiates mesenchymal stem cell survival via NF-κB-dependent miR-146a expression by targeting Fas
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DOI：
10.1152/ajpheart.00212.2010
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发表时间：
2010-10-01
影响因子：
4.8
通讯作者：
Haider, Husnain Kh
中科院分区：
文献类型：
--
作者：
Suzuki, Yoko;Kim, Ha Won;Haider, Husnain Kh
铃木 Y、金 HW、阿什拉夫 M、海德尔 HKh。二氮嗪通过靶向 Fas 的 NF-kappa B 依赖性 miR-146a 表达增强间充质干细胞的存活。 Am J Physiol Heart Circ Physiol 299：H1077-H1082，2010。首次发表于 2010 年 7 月 23 日； doi:10.1152/ajpheart.00212.2010.-我们之前曾报道过，用二氮嗪 (DZ) 预处理骨髓间充质干细胞 (MSC)，可通过 NF-kappa B 信号传导显着提高细胞存活率。由于 micro-RNA (miRNA) 是细胞凋亡、增殖和分化等多种生物事件的关键调节因子，因此 DZ 诱导的生存很可能是由 miRNA 介导的。在这里，我们表明，在 DZ 预处理过程中表达的 miR-146a 是干细胞存活的关键调节因子。通过乳酸脱氢酶释放和转移酶介导的 dUTP 缺口末端标记染色评估，用 DZ (200 μM) 处理 MSC 显着增加 miR-146a 表达并促进细胞存活。有趣的是，用 IKK-γ NEMO 结合域抑制剂肽阻断 NF-κ B 不会诱导 miR-146a 表达，表明 NF-κ B 调节 miR-146a 表达。此外，反义 miR-146a 抑制剂阻断 miR-146a 表达消除了 DZ 诱导的细胞保护作用，表明 miR-146a 在 MSC 存活中发挥关键作用。计算分析发现，Fas mRNA 3' 非翻译区中存在与细胞凋亡相关的 miR-146a 一致推定靶位点。 Fas 作为靶基因的作用通过 miR-146a 的废除得到证实，这显着增加了 Fas 蛋白的表达。这通过荧光素酶报告基因检测得到验证，结果表明 miR-146a 的强制表达通过靶向该基因的 3'-UTR 来下调 Fas 表达。综上所述，这些数据表明，用 DZ 预处理 MSC 所提供的细胞保护作用是通过 miR-146a 诱导来调节的，这可能是心脏缺血性疾病的新治疗靶点。
Suzuki Y, Kim HW, Ashraf M, Haider HKh. Diazoxide potentiates mesenchymal stem cell survival via NF-kappa B-dependent miR-146a expression by targeting Fas. Am J Physiol Heart Circ Physiol 299: H1077-H1082, 2010. First published July 23, 2010; doi:10.1152/ajpheart.00212.2010.-We have previously reported that preconditioning of bone marrow-derived mesenchymal stem cells (MSCs) with diazoxide (DZ) significantly improved cell survival via NF-kappa B signaling. Since micro-RNAs (miRNAs) are critical regulators of a wide variety of biological events, including apoptosis, proliferation, and differentiation, it is likely that DZ-induced survival is mediated by miRNAs. Here we show that miR-146a expressed during preconditioning with DZ is a key regulator of stem cell survival. Treatment of MSCs with DZ (200 mu M) markedly increased miR-146a expression and promoted cell survival, as evaluated by lactate dehydrogenase release and transferase-mediated dUTP nick-end labeling staining. Interestingly, blocking NF-kappa B by IKK-gamma NEMO binding domain inhibitor peptide did not induce miR-146a expression, indicating NF-kappa B regulates miR-146a expression. Moreover, blockade of miR-146a expression by antisense miR-146a inhibitor abolished DZ-induced cytoprotective effects, suggesting a critical role of miR-146a in MSC survival. Computational analysis found a consensus putative target site of miR-146a relevant to apoptosis in the 3' untranslated region of Fas mRNA. The role of Fas as a target gene was substantiated by abrogation of miR-146a, which markedly increased Fas protein expression. This was verified by luciferase reporter assay, which showed that forced expression of miR-146a downregulated Fas expression via targeting its 3'-UTR of this gene. Taken together, these data demonstrated that cytoprotection afforded by preconditioning of MSCs with DZ was regulated by miR-146a induction, which may be a novel therapeutic target in cardiac ischemic diseases.