MiR-21-5p in Macrophage-Derived Exosomes Targets Smad7 to Promote Epithelial Mesenchymal Transition of Airway Epithelial Cells.
MiR-21-5p in Macrophage-Derived Exosomes Targets Smad7 to Promote Epithelial Mesenchymal Transition of Airway Epithelial Cells.
复制标题
巨噬细胞源性外泌体中的 MiR-21-5p 靶向 Smad7 促进气道上皮细胞的上皮间质转化
DOI：
10.2147/jaa.s307165
复制
发表时间：
2021
影响因子：
3.2
通讯作者：
Shang Y
中科院分区：
文献类型：
--
作者：
Li X;Yang N;Cheng Q;Zhang H;Liu F;Shang Y
背景哮喘通常与气道炎症和气道重塑有关。上皮间质转化（EMT）常发生在气道重塑中。本研究旨在探讨巨噬细胞来源的exosomes中miR-21- 5 p和Smad 7信号通路对气道上皮细胞EMT的影响。方法采用HE染色和Masson染色法验证哮喘模型的成功建立。Western blot检测上皮细胞粘附因子和基质细胞标志物的表达。采用qRT-PCR检测miR-21- 5 p的表达水平。荧光原位杂交（FISH）进一步验证肺组织中miR-21- 5 p的表达。通过透射电子显微镜（TEM）和纳米颗粒跟踪分析（NTA）观察外泌体形态。采用荧光素酶报告基因分析miR-21- 5 p与Smad 7的相互作用。结果哮喘大鼠体内巨噬细胞中miR-21- 5 p表达上调。体外培养的肺泡巨噬细胞在LPS刺激下可分泌高水平miR-21- 5 p的exosomes。外泌体来源的miR-21- 5 p通过下调Smad 7，通过TGFβ1/Smad信号通路促进大鼠气管上皮细胞EMT。这一过程可被miR-21- 5 p抑制剂阻断。结论大鼠肺泡巨噬细胞产生高水平的含miR-21- 5 p的exosomes，将miR-21- 5 p转运至气管上皮细胞，通过TGF-β1/Smad信号通路靶向Smad 7促进EMT。
Background Asthma is usually associated with airway inflammation and airway remodeling. Epithelial mesenchymal transition (EMT) often occurs in airway remodeling. The purpose of this study is to identify the effect of miR-21-5p and Smad7 signaling pathway in macrophage-derived exosomes on EMT of airway epithelial cells. Methods HE staining and Masson staining were used to verify the successful establishment of the asthma model. The levels of epithelial cell adhesion factor and stromal cell markers were detected by Western blot. The levels of miR-21-5p were detected by qRT-PCR. The expression of miR-21-5p in lung tissue was further verified by fluorescence in situ hybridization (FISH). Exosome morphology was observed by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Luciferase reporter assay was applied to analyze the interaction of miR-21-5p with Smad7. Results The expression of miR-21-5p was upregulated in macrophages of rats in vivo with OVA-induced asthma. In vitro cultured alveolar macrophages stimulated by LPS could secrete exosomes with high levels of miR-21-5p. The exosome-derived miR-21-5p promotes EMT in rat tracheal epithelial cells through TGFβ1/Smad signaling pathway by downregulating Smad7. This process can be blocked by miR-21-5p inhibitor. Conclusion Rat alveolar macrophages produced high levels of miR-21-5p-containing exosomes, which transported miR-21-5p to tracheal epithelial cells, thus promoting EMT through TGF-β1/Smad signaling pathway by targeting Smad7.