MicroRNA-21 promotes wound healing via the Smad7-Smad2/3-Elastin pathway
MicroRNA-21 promotes wound healing via the Smad7-Smad2/3-Elastin pathway
复制标题
MicroRNA-21 通过 Smad7-Smad2/3-Elastin 途径促进伤口愈合
DOI：
10.1016/j.yexcr.2017.11.019
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发表时间：
2018-01-15
影响因子：
3.7
通讯作者：
Liu, Yi
中科院分区：
文献类型：
--
作者：
Li, Xiaoyan;Guo, Lijia;Liu, Yi
伤口愈合受到细胞、分子、细胞因子以及 microRNA (miRNA) 的复杂网络的调节。 miRNA被证实影响伤口愈合过程，而miRNA家族的重要成员miR-21也被证明可以调节伤口愈合。本研究的目的是研究 miR-21 在伤口愈合过程中的作用以及可能的潜在细胞信号通路。我们从 WT 和 miR-21-KO 小鼠牙龈中分离出 GMSC。使用流式细胞术分析和免疫细胞荧光染色来鉴定从 WT 和 miR-21-KO 小鼠获得的 GMSC。采用RT-PCR、蛋白质印迹分析和免疫组织荧光染色来检测细胞外基质成分和细胞信号通路关键蛋白的表达。 TargetScan 和 pmiR-RB-REPORT 载体用于验证 Smad7 是 miR-21 的直接靶标。与 WT 小鼠相比，miR-21-KO 小鼠的伤口愈合速度较慢。 RT-PCR 和蛋白质印迹分析表明，miR-21 缺陷样品中弹性蛋白表达下调。我们证实 Smad7 是 miR-21 的直接靶标。 miR-21 敲除导致 Smad7 表达增加，Smad2/3 复合物磷酸化受损。从WT和miR-21-KO小鼠获得的上颚组织切片中Smad7-Smad2/3-Elastin轴的表达显示出相同的趋势。基于所有这些结果，我们证明 miR-21 通过 Smad7-Smad2/3-Elastin 途径促进伤口愈合过程。
Wound healing is regulated by a complex network of cells, molecules, and cytokines, as well as microRNAs (miRNAs). miRNAs were confirmed to influence the wound healing process, and miR-21, an important member of the miRNA family, was also shown to regulate wound healing. The aim of the present study was to investigate the role of miR-21 in the wound healing process and the possible underlying cell signaling pathways. We isolated GMSCs from WT and miR-21-KO mouse gingiva. Flow cytometric analysis and immunocytofluorescense staining were used to identify the GMSCs acquired from WT and miR-21-KO mice. RT-PCR, western blot analysis and immunohistofluorescence staining were performed to examine the expression of extracellular matrix components and key proteins of cell signaling pathways. TargetScan and pmiR-RB-REPORT vectors were used to verify that Smad7 was a direct target of miR-21. Compared to WT mice, miR-21-KO mice showed slower wound healing. RT-PCR and western blot analysis indicated that Elastin expression was downregulated in miR-21-deficient samples. We confirmed that Smad7 was a direct target of miR-21. miR-21 knockout resulted in increased expression of Smad7 and impaired phosphorylation of the Smad2/3 complex. The expression of the Smad7-Smad2/3-Elastin axis in palate tissues sections acquired from WT and miR-21-KO mice showed the same trend. Based on all these results, we demonstrated that miR-21 promoted the wound healing process via the Smad7-Smad2/3-Elastin pathway.