Yan Bin, Qiu Jianing, Yang Yan, Luo Jing, Liu Dan, Zhang Lusi, Meng Zhishang, Li Huiling, Guo Xiaojian, Zeng Jun, Ding Chun, Xie Manyun, Long Kejun
Department of Ophthalmology, the Second Xiangya Hospital, Central South University, Changsha, China.
Hunan Clinical Research Center of Ophthalmic Disease, Changsha, China.
Invest Ophthalmol Vis Sci. 2025 Jul 1;66(9):24. doi: 10.1167/iovs.66.9.24.
Proliferative diabetic retinopathy (PDR) is a severe microvascular complication of diabetes mellitus. Although exosomes participate in various pathologic processes, their role in ocular pathologies is unclear. This study profiled exosomal microRNAs (exo-miRNAs) in the vitreous humor (VH) of patients with PDR to elucidate their regulatory roles in PDR pathogenesis.
VH samples were obtained from 23 patients with PDR and 17 non-diabetic controls. Exosomes were isolated using ultracentrifugation and characterized via transmission electron microscopy, nanoparticle tracking analysis and Western blot. High-throughput sequencing identified differentially expressed miRNAs (DEMs), followed by target gene prediction and bioinformatic analyses. The top DEMs were validated using quantitative real-time polymerase chain reaction (qRT-PCR).
Exosomes were isolated and characterized from VH samples. Sequencing analysis identified 843 unique miRNAs, with 60 showing significant differential expression between PDR and control groups (28 upregulated and 32 downregulated). The miR-451a and miR-486-5p were most upregulated; miR-204-5p and miR-211-5p were most downregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed DEM target genes enriched in endocrine, metabolic, signaling transduction, and intercellular adhesion pathways, notably cGMP-PKG, mTOR, and cAMP pathways. Network analysis identified miR-486-5p and miR-451a as key hubs. Validation by qRT-PCR confirmed the differential expression in the sequencing analysis.
This study provides the comprehensive profile of vitreous exo-miRNAs in patients with PDR, identifying key DEMs and their potential regulatory networks. These findings indicate that vitreous exo-miRNAs, specifically miR-204-5p and miR-486-5p, represent promising diagnostic biomarkers and potential therapeutic targets for PDR.
增殖性糖尿病视网膜病变（PDR）是糖尿病严重的微血管并发症。尽管外泌体参与多种病理过程，但其在眼部疾病中的作用尚不清楚。本研究分析了PDR患者玻璃体液（VH）中的外泌体微小RNA（exo-miRNA），以阐明其在PDR发病机制中的调控作用。
从23例PDR患者和17例非糖尿病对照者中获取VH样本。采用超速离心法分离外泌体，并通过透射电子显微镜、纳米颗粒跟踪分析和蛋白质印迹法进行鉴定。高通量测序鉴定差异表达的miRNA（DEM），随后进行靶基因预测和生物信息学分析。通过定量实时聚合酶链反应（qRT-PCR）验证 top DEM。
从VH样本中分离并鉴定了外泌体。测序分析鉴定出8个43种独特的miRNA，其中60种在PDR组和对照组之间表现出显著差异表达（28种上调，32种下调）。miR-451a和miR-486-5p上调最为明显；miR-204-5p和miR-211-5p下调最为明显。基因本体论和京都基因与基因组百科全书分析显示，DEM靶基因富集于内分泌、代谢、信号转导和细胞间粘附途径，尤其是cGMP-PKG、mTOR和cAMP途径。网络分析确定miR-486-5p和miR-451a为关键节点。qRT-PCR验证证实了测序分析中的差异表达。
本研究提供了PDR患者玻璃体外泌体miRNA的综合概况，确定了关键的DEM及其潜在调控网络。这些发现表明，玻璃体外泌体miRNA，特别是miR-204-5p和miR-486-5p，是PDR有前景的诊断生物标志物和潜在治疗靶点。
