Exosomes derived from human amniotic mesenchymal stem cells promotes angiogenesis in hUVECs by delivering novel miRNA N-194.

IF 6 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Medicine Pub Date : 2025-05-06 DOI:10.1186/s10020-025-01192-8

Yang Song, Tao Zhang, Ping Shi, Yingzhuo Gao, Xining Pang

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引用次数: 0

Abstract

Background: To investigate the effect and mechanism of exosomes derived from human amniotic mesenchymal stem cells (hAMSC-Exos) promoting angiogenesis.

Methods: HAMSC-Exos were isolated using ultracentrifugation and characterized by transmission electron microscopy, NTA, and Western blot. The uptake of hAMSC-Exos by hUVECs was analyzed using PKH-26 labeling, and the effect of hAMSC-Exos on angiogenesis was analyzed in human umbilical vein endothelial cells hUVECs by cell viability assay, Transwell migration assay, Matrigel tube formation assay, and Matrigel plug assays in nude mice. Bioinformatics methods were used to analyze miRNA high-throughput sequencing data of hAMSC-Exos, and RT-qPCR was used to validate the novel miRNAs. HAMSC-Exos with high and low N-194 expression were obtained by transfection, respectively. Target genes were predicted using TargetScan, and the mRNA and protein levels of potential target genes were analyzed by RT-qPCR and Western blot after N-194 mimics transfection. Interaction between miRNAs and target genes was detected using the dual-luciferase reporter assay. Target genes were overexpressed in hUVECs by transfection. The roles of target genes in the influence of N-194 on cell function were determined by analyzing angiogenesis.

Results: The extracted hAMSC-Exos showed saucer-shaped under transmission electron microscopy, and the NTA results showed the particle size of 115.6 ± 38.6 nm. The positive expression of CD9, CD63, and CD81 were verified using Western blot. The treatment of hUVECs with hAMSC-Exos significantly increased cell proliferation, migration, and angiogenesis. HAMSC-Exos contained the novel miRNAs N-194, N-314, N-19, N-393, and N-481, and the expression of N-194 was higher. The Exos derived from hAMSCs which were transfected with FAM-N-194 mimics were able to deliver FAM-N-194 mimics to hUVECs. The hAMSC-Exos with high N-194 significantly promoted angiogenesis in hUVECs. N-194 mimics transfection significantly reduced mRNA and protein levels of potential target gene ING5, and N-194 mimics significantly reduced the luciferase activities expressed by wild-type reporter gene vectors for ING5. The ING5 overexpression significantly reduced the angiogenic capacity of hUVECs. ING5 overexpression suppressed the expression of HSP27 and PLCG2.

Conclusions: HAMSC-Exos promotes angiogenesis in hUVECs by delivering novel miRNA N-194 which targets ING5.

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来自人羊膜间充质干细胞的外泌体通过传递新的miRNA N-194促进huvec的血管生成。

背景：探讨人羊膜间充质干细胞（hAMSC-Exos）外泌体促进血管生成的作用及其机制。方法：采用超离心分离HAMSC-Exos，采用透射电镜、NTA和Western blot对其进行鉴定。采用ph -26标记法分析hAMSC-Exos对人脐静脉内皮细胞hUVECs的摄取情况，并采用细胞活力法、Transwell迁移法、Matrigel管形成法、Matrigel塞法分析hAMSC-Exos对人脐静脉内皮细胞血管生成的影响。采用生物信息学方法对hAMSC-Exos的miRNA高通量测序数据进行分析，并采用RT-qPCR对新miRNA进行验证。分别转染得到N-194高表达和低表达的HAMSC-Exos。应用TargetScan预测靶基因，转染N-194模拟物后，应用RT-qPCR和Western blot分析潜在靶基因的mRNA和蛋白水平。使用双荧光素酶报告基因法检测mirna与靶基因之间的相互作用。转染hUVECs后，靶基因过表达。通过血管生成分析，确定靶基因在N-194对细胞功能影响中的作用。结果：提取的hAMSC-Exos在透射电镜下呈碟状，NTA结果显示粒径为115.6±38.6 nm。Western blot检测CD9、CD63、CD81的阳性表达。用hAMSC-Exos治疗hUVECs可显著增加细胞增殖、迁移和血管生成。HAMSC-Exos中含有N-194、N-314、N-19、N-393和N-481新miRNAs，其中N-194的表达量较高。用FAM-N-194模拟物转染的hAMSCs衍生的Exos能够将FAM-N-194模拟物传递到hUVECs。高N-194的hAMSC-Exos显著促进hUVECs血管生成。N-194模拟物转染显著降低了潜在靶基因ING5的mRNA和蛋白水平，并且N-194模拟物显著降低了野生型ING5报告基因载体表达的荧光素酶活性。ING5过表达显著降低hUVECs的血管生成能力。ING5过表达可抑制HSP27和PLCG2的表达。结论：HAMSC-Exos通过传递靶向ING5的新型miRNA N-194促进huvec血管生成。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Medicine 医学-生化与分子生物学

CiteScore

8.60

自引率

0.00%

发文量

137

审稿时长

1 months

期刊介绍： Molecular Medicine is an open access journal that focuses on publishing recent findings related to disease pathogenesis at the molecular or physiological level. These insights can potentially contribute to the development of specific tools for disease diagnosis, treatment, or prevention. The journal considers manuscripts that present material pertinent to the genetic, molecular, or cellular underpinnings of critical physiological or disease processes. Submissions to Molecular Medicine are expected to elucidate the broader implications of the research findings for human disease and medicine in a manner that is accessible to a wide audience.