Selective microRNA expression of exosomes from retinal pigment epithelial cells by oxidative stress

IF 1.5 4区 心理学 Q4 NEUROSCIENCES Vision Research Pub Date : 2024-04-08 DOI:10.1016/j.visres.2024.108388
Zhengyu Zhang , Qinyuan Gu , Lu Chen , Dongqing Yuan , Xunyi Gu , Huiming Qian , Ping Xie , Qinghuai Liu , Zizhong Hu
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Abstract

The function of exosomal miRNAs (miRs) in retinal degeneration is largely unclear. We were aimed to investigate the functions of exosomes as well as their miRs derived from retinal pigment epithelial (RPE) cells following exposure to oxidative stress (OS). After the OS by lipopolysaccharide and rotenone on RPE cells, interleukin-1 beta (IL-1β), Interleukin-6 (IL-6), Tumor Necrosis Factor-alpha (TNF-α) were upregulated, along with the decreased mitochondrial membrane potential and upregulated oxidative damage marker 8-OH-dG in RPE cells. RPE-derived exosomes were then isolated, identified, injected into the subretinal space in mice. After subretinal injection, RPE-exosomes after OS not only induced higher ROS level and apoptotic retinal cells, but also elevated IL-1β, IL-6 alongside TNF-α expressions among retina/RPE/choroidal complex. Next, miRs inside the exosomes were sequenced by the next generation sequencing (NGS) technology. NGS revealed that certain miRs were abundant in exosomes, while others were selectively kept by RPE cells. Further, downregulated miRs, like miR-125b-5p, miR-125a-5p, alongside miR-128-3p, and upregulated miR, such as miR-7-5p were validated byRT-qPCR. Finally, Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were used to find the possible target genes of those selective exosomal miRs. Our results proved that the RPE-derived exosomes after OS selectively express certain miRs, providing novel insights into the pathogenesis of age-related macular degeneration (AMD) in future.

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氧化应激对视网膜色素上皮细胞外泌体微RNA表达的选择性影响
外泌体 miRNA(miRs)在视网膜变性中的功能尚不清楚。我们的目的是研究视网膜色素上皮细胞(RPE)暴露于氧化应激(OS)后产生的外泌体及其 miRs 的功能。RPE细胞受到脂多糖和鱼藤酮氧化应激后,白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)上调,线粒体膜电位下降,氧化损伤标志物8-OH-dG上调。然后分离、鉴定 RPE 衍生的外泌体,并将其注入小鼠视网膜下间隙。视网膜下注射后,OS后的RPE外泌体不仅诱导了更高的ROS水平和视网膜细胞凋亡,还升高了视网膜/RPE/脉络膜复合体中IL-1β、IL-6和TNF-α的表达。接着,利用新一代测序(NGS)技术对外泌体中的miRs进行了测序。NGS 发现,某些 miRs 在外泌体中含量丰富,而另一些则被 RPE 细胞选择性地保留下来。此外,下调的 miR(如 miR-125b-5p、miR-125a-5p 和 miR-128-3p)和上调的 miR(如 miR-7-5p)也通过 RT-qPCR 得到了验证。最后,通过基因本体(GO)注释和京都基因组百科全书(KEGG)通路富集分析,找到了这些选择性外泌体 miRs 的可能靶基因。我们的研究结果证明,OS后RPE衍生的外泌体选择性地表达了某些miRs,这为今后研究老年性黄斑变性(AMD)的发病机制提供了新的视角。
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来源期刊
Vision Research
Vision Research 医学-神经科学
CiteScore
3.70
自引率
16.70%
发文量
111
审稿时长
66 days
期刊介绍: Vision Research is a journal devoted to the functional aspects of human, vertebrate and invertebrate vision and publishes experimental and observational studies, reviews, and theoretical and computational analyses. Vision Research also publishes clinical studies relevant to normal visual function and basic research relevant to visual dysfunction or its clinical investigation. Functional aspects of vision is interpreted broadly, ranging from molecular and cellular function to perception and behavior. Detailed descriptions are encouraged but enough introductory background should be included for non-specialists. Theoretical and computational papers should give a sense of order to the facts or point to new verifiable observations. Papers dealing with questions in the history of vision science should stress the development of ideas in the field.
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