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评估ARPE-19细胞中微小RNA对氧化应激的反应。

Evaluation of microRNA responses in ARPE-19 cells against the oxidative stress.

作者信息

Ayaz Lokman, Dinç Erdem

机构信息

a Department of Biochemistry , School of Pharmacy, Trakya University , Edirne , Turkey.

b Department of Ophthalmology , School of Medicine, Mersin University , Mersin , Turkey.

出版信息

Cutan Ocul Toxicol. 2018 Jun;37(2):121-126. doi: 10.1080/15569527.2017.1355314. Epub 2017 Jul 31.

Abstract

PURPOSE

This study aimed to determine microRNA (miRNA) expression profile of human retinal pigment epithelium cell (ARPE-19) against the oxidative stress induced by hydrogen peroxide (HO).

METHODS

ARPE-19 cells were incubated with different concentrations of HO (200, 600 and 800 μM) for 18 h, and then cell viability, vascular endothelial growth factor levels and total oxidant status were evaluated. Expressions of 1152 miRNA were determined by quantitative real-time PCR in each group.

RESULTS

Expressions of 90 miRNA were significantly changed in the ARPE-19 cells incubated with HO compared to control group. However, miR-143-3p was only found to be expressed in groups incubated with HO. While 24 miRNA (hsa-miR-200c-3p, miR-192-5p, miR-194-5p, miR-141-3p, miR-658, miR-18 b-5p, miR-486-5p, miR-525-3p, miR-493-3p, miR-518d-3p, miR-29 b-1-5p, miR-675-3p, miR-1238-3p, miR-195-3p, miR-1539, miR-490-5p, miR-3200-5p, miR-1273d, miR-130a-5p, miR-30 b-5p, miR-1247-5p, miR-1910-5p, miR27a-5p and miR-200 b-3p) upregulated due to the increased dose of HO, nine miRNA (hsa-miR-96-5p, miR-33a-5p, miR-345-5p, miR-106 b-3p, miR-1285-3p, miR-23 b-5p, miR-27 b-5p, miR-103a-3p and miR-4289) were also found to be downregulated.

CONCLUSION

This study suggests that oxidative stress may be an important factor on expression of miRNAs in ARPE-19 cells. These miRNAs may have a role in the pathogenesis of age-related macular degeneration related to oxidative stress. However, this relationship needs to be examined in new studies by evaluation of pathways and target genes.

摘要

目的

本研究旨在确定人视网膜色素上皮细胞(ARPE - 19)在过氧化氢(H₂O₂)诱导的氧化应激下的微小RNA(miRNA)表达谱。

方法

将ARPE - 19细胞与不同浓度的H₂O₂(200、600和800μM)孵育18小时,然后评估细胞活力、血管内皮生长因子水平和总氧化剂状态。通过定量实时PCR测定每组中1152种miRNA的表达。

结果

与对照组相比,在与H₂O₂孵育的ARPE - 19细胞中,90种miRNA的表达发生了显著变化。然而,仅在与H₂O₂孵育的组中发现miR - 143 - 3p表达。随着H₂O₂剂量增加,24种miRNA(hsa - miR - 200c - 3p、miR - 192 - 5p、miR - 194 - 5p、miR - 141 - 3p、miR - 658、miR - 18b - 5p、miR - 486 - 5p、miR - 525 - 3p、miR - 493 - 3p、miR - 518d - 3p、miR - 29b - 1 - 5p、miR - 675 - 3p、miR - 1238 - 3p、miR - 195 - 3p、miR - 1539、miR - 490 - 5p、miR - 3200 - 5p、miR - 1273d、miR - 130a - 5p、miR - 30b - 5p、miR - 1247 - 5p、miR - 1910 - 5p、miR27a - 5p和miR - 200b - 3p)上调,另外9种miRNA(hsa - miR - 96 - 5p、miR - 33a - 5p、miR - 345 - 5p、miR - 106b - 3p、miR - 1285 - 3p、miR - 23b - 5p、miR - 27b - 5p、miR - 103a - 3p和miR - 4289)下调。

结论

本研究表明氧化应激可能是ARPE - 19细胞中miRNA表达的重要因素。这些miRNA可能在与氧化应激相关的年龄相关性黄斑变性的发病机制中起作用。然而,这种关系需要在新的研究中通过评估相关途径和靶基因来进一步研究。

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