衰老内皮细胞中功能性小细胞外囊泡的产生增加。

IF 5.3 2区 医学 Q1 Biochemistry, Genetics and Molecular Biology
Journal of Cellular and Molecular Medicine Pub Date : 2020-04-01 Epub Date: 2020-02-26 DOI:10.1111/jcmm.15047
Jaime A Riquelme, Kaloyan Takov, Concepción Santiago-Fernández, Xavier Rossello, Sergio Lavandero, Derek M Yellon, Sean M Davidson
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引用次数: 30

摘要

小细胞外囊泡(EVs)是血管生物学中的新成员。然而,对它们的产生和功能的彻底了解仍然是难以捉摸的。内皮细胞衰老是血管老化的一个关键特征,因此是治疗血管疾病的一个有吸引力的治疗靶点。在这项研究中,我们试图表征衰老内皮细胞的EV生产。为了实现这一目标,通过显微镜和流式细胞术测量衰老相关β-半乳糖苷酶活性,人类脐血管内皮细胞(HUVECs)被复制,直到它们达到衰老。免疫荧光法检测内体标记物Rab7和EV标记物CD63的表达。通过超离心分离小ev,利用电镜、纳米颗粒跟踪分析和免疫分析对其进行表征,评估外泌体标志物CD9和CD81的形态、大小、浓度和表达。利用transwell实验研究了HUVECs对ev的迁移反应。结果表明,衰老内皮细胞表达Rab7和CD63水平升高。此外,衰老的内皮细胞产生更高水平的CD9-和cd81阳性ev。此外,来自年轻和衰老内皮细胞的小ev促进了HUVEC的迁移。总的来说,衰老的内皮细胞产生更多的功能性小EVs,这可能在血管生理和疾病中起作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Increased production of functional small extracellular vesicles in senescent endothelial cells.

Increased production of functional small extracellular vesicles in senescent endothelial cells.

Increased production of functional small extracellular vesicles in senescent endothelial cells.

Small extracellular vesicles (EVs) are novel players in vascular biology. However, a thorough understanding of their production and function remains elusive. Endothelial senescence is a key feature of vascular ageing and thus, is an attractive therapeutic target for the treatment of vascular disease. In this study, we sought to characterize the EV production of senescent endothelial cells. To achieve this, Human Umbilical Vascular Endothelial Cells (HUVECs) were replicated until they reached senescence, as determined by measurement of Senescence-Associated β-Galactosidase activity via microscopy and flow cytometry. Expression of the endosomal marker Rab7 and the EV marker CD63 was determined by immunofluorescence. Small EVs were isolated by ultracentrifugation and characterized using electron microscopy, nanoparticle tracking analysis and immunoassays to assess morphology, size, concentration and expression of exosome markers CD9 and CD81. Migration of HUVECs in response to EVs was studied using a transwell assay. The results showed that senescent endothelial cells express higher levels of Rab7 and CD63. Moreover, senescent endothelial cells produced higher levels of CD9- and CD81-positive EVs. Additionally, small EVs from both young and senescent endothelial cells promoted HUVEC migration. Overall, senescent endothelial cells produce an increased number of functional small EVs, which may have a role in vascular physiology and disease.

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来源期刊
CiteScore
10.00
自引率
1.90%
发文量
496
审稿时长
28 weeks
期刊介绍: Bridging physiology and cellular medicine, and molecular biology and molecular therapeutics, Journal of Cellular and Molecular Medicine publishes basic research that furthers our understanding of the cellular and molecular mechanisms of disease and translational studies that convert this knowledge into therapeutic approaches.
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