泪膜 microRNAs 及其与人类干眼症的关系。
Tear Film miRNAs and Their Association With Human Dry Eye Disease.
机构信息
School of Optometry, Department of Optometry and Vision Science, University of Alabama at Birmingham.
出版信息
Curr Eye Res. 2022 Nov;47(11):1479-1487. doi: 10.1080/02713683.2022.2110597. Epub 2022 Aug 19.
PURPOSE
miRNAs can regulate inflammatory pathways. The purpose of this work was to determine if inflammatory-related tear film miRNAs are associated with extracellular vesicles (EVs) in human non-Sjögren's Syndrome dry eye disease (DED) participants.
METHODS
Five DED and 5 non-DED human participants were recruited. Tears samples were collected by washing the ocular surface of both eyes with phosphate buffered saline, pooling samples from the right and left eyes, and purifying EVs from the samples with a polyethylene glycol (PEG) 8000 precipitation procedure. Samples were directly analyzed via ELISA or transmission electron microscopy (TEM), or RNA was isolated first from the EVs and evaluated with RNA-Seq.
RESULTS
EVs were identified in the tear film of both groups using TEM and ELISA. Following EV purification and RNA isolation, RNA-Seq determined that there were 126 EV miRNAs differentially expressed between the two groups when comparing their RNA cargoes. Ingenuity Pathways Analysis found 9 upregulated miRNAs that were associated with inflammation (miR-127-5p, miR-1273h-3p, miR-1288-5p, miR-130b-5p, miR-139-3p, miR-1910-5p, miR-203b-5p, miR-22-5p, and miR-4632-3p; all < 0.049; fold regulation range = 1.43-1.67).
CONCLUSION
This study determined that EVs are present in the tear film and that tear EVs contain miRNAs that may be associated with DED inflammatory pathways.
目的
miRNA 可以调节炎症途径。本研究旨在确定炎症相关的泪膜 miRNA 是否与人类非干燥综合征干眼症（DED）患者的细胞外囊泡（EV）有关。
方法
招募了 5 名 DED 和 5 名非 DED 患者。通过用磷酸盐缓冲盐水冲洗双眼的眼表面收集泪液样本，将右眼和左眼的样本合并，并使用聚乙二醇（PEG）8000 沉淀程序从样本中纯化 EV。通过 ELISA 或透射电子显微镜（TEM）直接分析样品，或首先从 EV 中分离 RNA 并进行 RNA-Seq 评估。
结果
使用 TEM 和 ELISA 在两组泪膜中均鉴定出 EV。在 EV 纯化和 RNA 分离后，RNA-Seq 确定当比较两组的 RNA 载量时，有 126 个 EV miRNA 在两组之间存在差异表达。通路分析发现 9 个上调的 miRNA 与炎症相关（miR-127-5p、miR-1273h-3p、miR-1288-5p、miR-130b-5p、miR-139-3p、miR-1910-5p、miR-203b-5p、miR-22-5p 和 miR-4632-3p；均<0.049；倍数调控范围=1.43-1.67）。
结论
本研究确定 EV 存在于泪膜中，并且泪 EV 中可能含有与 DED 炎症途径相关的 miRNA。
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