| miR-140-5p靶向Nrf2对高糖诱导的视网膜色素上皮细胞氧化应激的调节作用 |
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| 引用本文： | 张新霞,陆丽红,狄文玉,王小敏,王保君. miR-140-5p靶向Nrf2对高糖诱导的视网膜色素上皮细胞氧化应激的调节作用[J]. 眼科新进展, 2020, 0(8): 727-730. DOI: 10.13389/j.cnki.rao.2020.0165 |
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| 作者姓名： | 张新霞 陆丽红 狄文玉 王小敏 王保君 |
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| 作者单位： | 453100 河南省新乡市,新乡医学院第一附属医院眼科（张新霞，陆丽红，王小敏，王保君）；453100 河南省新乡市,新乡医学院第一附属医院病理科（狄文玉） |
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| 基金项目： | 河南省医学科技攻关计划项目(编号:SBGJ2018055)； |
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| 摘 要： | 目的探讨miR-140-5p靶向核因子-类胡萝卜素2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)对高糖诱导的视网膜色素上皮(retinal pigment epithelial,RPE)细胞氧化应激的调节作用及对应的分子生物学机制。方法使用5 mmol·L-1、10 mmol·L-1、20 mmol·L-1、30 mmol·L-1、50mmol·L-1的葡萄糖分别处理ARPE-19细胞。利用CCK-8法及RT-qPCR检测各葡萄糖浓度下ARPE-19细胞的活力及miR-140-5p、Nrf2 mRNA表达情况。将miR-140-5p inhibitor、miR-140-5p-NC、miR-140-5p-mimic、si-Con或si-Nrf2转染入ARPE-19细胞,根据转染物和葡萄糖浓度不同分组,使用DCFH-DA荧光探针检测各组细胞内活性氧(reactive oxygen species,ROS)水平,使用超...
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| 关 键 词： | miR-140-5p 核因子-类胡萝卜素2相关因子2 高糖 视网膜色素上皮细胞 氧化应激 |
miRNA-140-5p modulates high glucose-induced oxidative stress of retinal pigment epithelial cells by targeting Nrf2 |
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| ZHANG Xinxia1,LU Lihong1,DI Wenyu2,WANG Xiaomin1,WANG Baojun1. miRNA-140-5p modulates high glucose-induced oxidative stress of retinal pigment epithelial cells by targeting Nrf2[J]. Recent Advances in Ophthalmology, 2020, 0(8): 727-730. DOI: 10.13389/j.cnki.rao.2020.0165 |
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| Authors: | ZHANG Xinxia1 LU Lihong1 DI Wenyu2 WANG Xiaomin1 WANG Baojun1 |
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| Affiliation: | 1.Department of Ophthalmology，the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100, Henan Province, China2.Department of Pathology，the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100, Henan Province, China |
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| Abstract: | Objective To investigate the molecular mechanism of oxidative stress induced by miR-140-5p in retinal pigment epithelial cells (RPE) induced by high glucose (HG) by targeting nuclear factor erythroid 2-related factor 2 (Nrf2).Methods ARPE-19 cells were treated with glucose at concentrations of 5 mmol·L-1, 10 mmol·L-1, 20 mmol·L-1, 30 mmol·L-1, 50 mmol·L-1 respectively. CCK-8 was used to detect the survival rate and RT-qPCR was applied to detect miR-140-5p mRNA expression of RPE cells with increasing glucose concentration in turn; DCFH-DA fluorescence probe was used to detect the level of reactive oxygen species (ROS) in cells treated with interfering miR-140-5p, MDA and SOD enzyme activity kit were used to detect superoxide dismutase (SOD) activities, Nrf2 mRNA and protein expression were detected by RT-PCR and Western blot, and the target was verified by luciferase reporter gene experiment.Results Compared with the low glucose condition of 5 mmol·L-1, the survival rate of RPE cells of 30 mmol·L-1 and 50 mmol·L-1 decreased significantly (P<0.05, P<0.001) and the expression level of miR-140-5p increased significantly (P<0.05, P<0.001). Compared with the 50 mmol·L-1 glucose control group, the transfection of miR-140-5p inhibitor significantly increased cell survival rate, decreased ROS content and increased SOD activity (all P<0.01). Compared with the low glucose conditions of 5 mmol·L-1, the Nrf2 mRNA expression levels of RPE cells of 30 mmol·L-1 and 50 mmol·L-1 decreased significantly (P<0.01, P<0.001). The results of luciferase test showed that, compared with the empty control group, the transfection of miR-140-5p mimic significantly decreased the Nrf2 activity, whereas the transfection of miR-140-5p inhibitor significantly increased the Nrf2 activity, with significant difference (both P<0.001). At 50 mmol·L-1 glucose concentration, compared with the RPE cells transfected with miR-140-5p-inhibitor+si-Con, the RPE cells transfected with miR-140-5p inhibitor and si-Nrf2 significantly decreased cell survival rate, decreased SOD activity, and increased ROS content (all P<0.01).Conclusion miR-140-5p can modulate high glucose-induced oxidative stress injury of RPE cells by targeting Nrf2. |
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| Keywords: | miR-140-5p nuclear factor erythroid 2-related factor 2 high glucose retinal pigment epithelial cells oxidative stress |
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