Oxidative stress in retinal pigment epithelium cells increases exosome secretion and promotes angiogenesis in endothelial cells.
Oxidative stress in retinal pigment epithelium cells increases exosome secretion and promotes angiogenesis in endothelial cells.
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DOI：
10.1111/jcmm.12834
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发表时间：
2016-08
影响因子：
5.3
通讯作者：
Sancho-Pelluz J
中科院分区：
文献类型：
--
作者：
Atienzar-Aroca S;Flores-Bellver M;Serrano-Heras G;Martinez-Gil N;Barcia JM;Aparicio S;Perez-Cremades D;Garcia-Verdugo JM;Diaz-Llopis M;Romero FJ;Sancho-Pelluz J
视网膜色素上皮(RPE)是位于光感受器和脉络膜之间的单层，经常受到氧化应激的破坏，特别是由于活性氧(ROS)的作用。由于RPE的生理功能，对视网膜的生存是必不可少的，任何持续的损伤都可能导致视力丧失。Exosome是由多种细胞类型释放到细胞外介质中的小的膜性囊泡，包括RPE细胞。它们的货物包括遗传物质和蛋白质，这使得这些小泡对细胞间的交流至关重要。外体可能会与邻近的细胞融合，影响它们的命运。已经观察到，当RPE细胞处于氧化应激状态时，它们会释放更多的外切体。用超速离心法分离培养的RPE细胞的外切体，并用流式细胞仪进行定量。用流式细胞仪和Western印迹法检测血管内皮生长因子受体(VEGFR)的表达。采用逆转录聚合酶链式反应(RT-PCR)和定量聚合酶链式反应(QPCR)检测血管内皮生长因子受体(VEGFRs)的mRNA含量。将RPE外切体应用于内皮细胞培养后，进行新生血管分析。我们的结果显示，应激的RPE细胞释放了比对照更多的外切体，膜上VEGFR的表达更高，并包裹了额外的VEGFR mRNA。血管生成分析证实，当暴露于应激的RPE外切体时，内皮细胞增加了其管状形成能力。
The retinal pigment epithelium (RPE), a monolayer located between the photoreceptors and the choroid, is constantly damaged by oxidative stress, particularly because of reactive oxygen species (ROS). As the RPE, because of its physiological functions, is essential for the survival of the retina, any sustained damage may consequently lead to loss of vision. Exosomes are small membranous vesicles released into the extracellular medium by numerous cell types, including RPE cells. Their cargo includes genetic material and proteins, making these vesicles essential for cell‐to‐cell communication. Exosomes may fuse with neighbouring cells influencing their fate. It has been observed that RPE cells release higher amounts of exosomes when they are under oxidative stress. Exosomes derived from cultured RPE cells were isolated by ultracentrifugation and quantified by flow cytometry. VEGF receptors (VEGFR) were analysed by both flow cytometry and Western blot. RT‐PCR and qPCR were conducted to assess mRNA content of VEGFRs in exosomes. Neovascularization assays were performed after applying RPE exosomes into endothelial cell cultures. Our results showed that stressed RPE cells released a higher amount of exosomes than controls, with a higher expression of VEGFR in the membrane, and enclosed an extra cargo of VEGFR mRNA. Angiogenesis assays confirmed that endothelial cells increased their tube formation capacity when exposed to stressed RPE exosomes.