Exosomes derived from human amniotic epithelial cells accelerate diabetic wound healing via PI3K-AKT-mTOR-mediated promotion in angiogenesis and fibroblast function
Exosomes derived from human amniotic epithelial cells accelerate diabetic wound healing via PI3K-AKT-mTOR-mediated promotion in angiogenesis and fibroblast function
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源自人羊膜上皮细胞的外泌体通过 PI3K-AKT-mTOR 介导的血管生成和成纤维细胞功能促进加速糖尿病伤口愈合
DOI：
10.1093/burnst/tkaa020
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发表时间：
2020-01-01
期刊：
影响因子：
5.3
通讯作者：
Xia, Zhaofan
中科院分区：
文献类型：
--
作者：
Wei, Pei;Zhong, Chenjian;Xia, Zhaofan
背景糖尿病创面是糖尿病最常见、最严重的并发症之一，其特点是创面愈合相关细胞在数量和质量上出现功能障碍。我们之前的研究表明，人羊膜上皮细胞（hAECs）可以通过旁分泌作用促进糖尿病伤口愈合。有趣的是，许多研究表明，来自干细胞的外泌体是干细胞治疗的关键旁分泌载体。然而，来自hAECs的外泌体（hAECs- exos）是否介导hAECs对糖尿病伤口愈合的影响尚不清楚。本研究旨在探讨haec - exos对糖尿病创面愈合的生物学作用，并初步阐明其作用机制。方法采用超离心分离haec - exos，透射电镜、动态光散射和流式细胞术对其进行鉴定。通过一系列体外功能分析来评估haec - exos在高血糖微环境下对人成纤维细胞（HFBs）和人脐静脉内皮细胞（HUVECs）的调节作用。通过高通量测序和生物信息学分析，推测haec - exos对hbs和HUVECs作用的相关机制。随后，我们评估了候选信号通路haec - exos在调节huvec和HFBs功能以及糖尿病伤口愈合中的作用。结果haec - exos呈杯状或球形，平均直径为105.89±10.36 nm， CD63和TSG101阳性，可被hbs和HUVECs内化。之后，haec - exos不仅显著促进了hbs的增殖和迁移，还促进了HUVECs的体外血管生成活性。高通量测序显示haec - exos参与伤口愈合的mirna富集。京都基因与基因组百科全书和基因本体分析表明，前15个mirna的靶基因在PI3K-AKT通路中高度富集。进一步的功能研究表明，PI3K-AKT-mTOR通路对于haec - exos诱导糖尿病小鼠HFBs和huvec的生物学效应以及伤口愈合是必要的。我们的研究结果表明，haec - exos通过激活PI3K-AKT-mTOR通路促进血管生成和成纤维细胞功能，为糖尿病伤口愈合提供了一种有前景的新策略。
Abstract Background Diabetic wounds are one of the most common and serious complications of diabetes mellitus, characterized by the dysfunction of wound-healing-related cells in quantity and quality. Our previous studies revealed that human amniotic epithelial cells (hAECs) could promote diabetic wound healing by paracrine action. Interestingly, numerous studies demonstrated that exosomes derived from stem cells are the critical paracrine vehicles for stem cell therapy. However, whether exosomes derived from hAECs (hAECs-Exos) mediate the effects of hAECs on diabetic wound healing remains unclear. This study aimed to investigate the biological effects of hAECs-Exos on diabetic wound healing and preliminarily elucidate the underlying mechanism. Methods hAECs-Exos were isolated by ultracentrifugation and identified by transmission electron microscopy, dynamic light scattering and flow cytometry. A series of in vitro functional analyses were performed to assess the regulatory effects of hAECs-Exos on human fibroblasts (HFBs) and human umbilical vein endothelial cells (HUVECs) in a high-glycemic microenvironment. High-throughput sequencing and bioinformatics analyses were conducted to speculate the related mechanisms of actions of hAECs-Exos on HFBs and HUVECs. Subsequently, the role of the candidate signaling pathway of hAECs-Exos in regulating the function of HUVECs and HFBs, as well as in diabetic wound healing, was assessed. Results hAECs-Exos presented a cup- or sphere-shaped morphology with a mean diameter of 105.89 ± 10.36 nm, were positive for CD63 and TSG101 and could be internalized by HFBs and HUVECs. After that, hAECs-Exos not only significantly promoted the proliferation and migration of HFBs, but also facilitated the angiogenic activity of HUVECs in vitro. High-throughput sequencing revealed enriched miRNAs of hAECs-Exos involved in wound healing. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses have shown that the target genes of the top 15 miRNAs were highly enriched in the PI3K-AKT pathway. Further functional studies demonstrated that the PI3K-AKT-mTOR pathway was necessary for the induced biological effects of hAECs-Exos on HFBs and HUVECs, as well as on wound healing, in diabetic mice. Conclusions Our findings demonstrated that hAECs-Exos represent a promising, novel strategy for diabetic wound healing by promoting angiogenesis and fibroblast function via activation of the PI3K-AKT-mTOR pathway.