Exosomal let-7i-5p from three-dimensional cultured human umbilical cord mesenchymal stem cells inhibits fibroblast activation in silicosis through targeting TGFBR1
Exosomal let-7i-5p from three-dimensional cultured human umbilical cord mesenchymal stem cells inhibits fibroblast activation in silicosis through targeting TGFBR1
复制标题
三维培养人脐带间充质干细胞外泌体let-7i-5p通过靶向TGFBR1抑制矽肺成纤维细胞活化
DOI：
10.1016/j.ecoenv.2022.113302
复制
发表时间：
2022-02-18
影响因子：
6.8
通讯作者：
Tian, Lin
中科院分区：
文献类型：
--
作者：
Xu, Chunjie;Hou, Lin;Tian, Lin
矽肺肺纤维化（PF）与长期过量吸入二氧化硅有关。成纤维细胞活化为肌成纤维细胞是导致肺纤维化的主要终末效应，这对研究矽肺纤维化的发生发展及其防治具有重要意义。来源于人脐带间充质干细胞的外泌体（hucMSC-Exos）被认为是二氧化硅诱导的PF的潜在治疗方法，然而其确切机制仍不清楚。因此，本研究旨在探讨hucMSC-Exos是否影响成纤维细胞的活化以缓解PF。本研究采用三维（3D）方法培养hucMSC和MRC-5细胞（人胚肺成纤维细胞），并从无血清培养基中分离外泌体，通过纳米颗粒追踪分析（NTA）、透射电子显微镜（TEM）和蛋白质印迹分析进行鉴定。然后，本研究使用二氧化硅诱导的PF的动物模型，观察hucMSC-Exos和MRC-5-Exos对成纤维细胞活化的影响。此外，在NIH-3 T3细胞（小鼠胚胎成纤维细胞）中通过蛋白质印迹分析、伤口愈合和用hucMSC-Exos和MRC-5-Exos处理的迁移测定来分析成纤维细胞的活化。此外，通过高通量测序测量了hucMSCs-Exos和MRC-5-Exos之间microRNA（DE miRNAs）的差异表达。HucMSC-Exos抑制小鼠和NIH-3 T3细胞中成纤维细胞的活化。与MRC-5-Exos相比，Let-7i-5 p在hucMSCs-Exos中显著上调，这与二氧化硅诱导的PF有关。hucMSCs-Exos的Let-7i-5 p负责通过靶向TGFBR 1激活成纤维细胞。同时，Smad 3在成纤维细胞的活化中也起重要作用。该研究表明，hucMSCs-Exos作为一种介导剂，转移let-7i-5 p以抑制成纤维细胞的活化，其通过TGFBR 1/Smad 3信号通路使PF活化。该机理对硅源性肺纤维化的治疗具有潜在的应用价值。
Silicosis of pulmonary fibrosis (PF) is related to long-term excessive inhalation of silica. The activation of fibroblasts into myofibroblasts is the main terminal effect leading to lung fibrosis, which is of great significance to the study of the occurrence and development of silicosis fibrosis and its prevention and treatment. Exosomes derived from human umbilical cord mesenchymal stem cells (hucMSC-Exos) are considered to be a potential therapy of silica-induced PF, however, their exact mechanism remains unknown. Therefore, this study aims to explore whether hucMSC-Exos affect the activation of fibroblasts to alleviate PF. In this study, a threedimensional (3D) method was applied to culture hucMSCs and MRC-5 cells (human embryonic lung fibroblasts), and exosomes were isolated from serum-free media, identified by nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM) and Western blotting analysis. Then, the study used an animal model of silica-induced PF to observe the effects of hucMSC-Exos and MRC-5-Exos on activation of fibroblasts. In addition, the activation of fibroblasts was analyzed by Western blotting analysis, wound healing, and migration assay with the treatment of hucMSC-Exos and MRC-5-Exos in NIH-3T3 cells (mouse embryonic fibroblasts). Furthermore, differential expression of microRNAs (DE miRNAs) was measured between hucMSCs-Exos and MRC-5-Exos by high throughput sequence. HucMSC-Exos inhibited the activation of fibroblasts in mice and NIH-3T3 cells. Let-7i5p was significantly up-regulated in hucMSCs-Exos compared to MRC-5-Exos, which was related to silica-induced PF. Let-7i-5p of hucMSCs-Exos was responsible for the activation of fibroblasts by targeting TGFBR1. Meanwhile, Smad3 was also an important role in the activation of fibroblasts. The study demonstrates that hucMSCs-Exos act as a mediator that transfers let-7i-5p to inhibit the activation of fibroblasts, which alleviates PF through the TGFBR1/Smad3 signaling pathway. The mechanism has potential value for the treatment of silica-induced PF.