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蓝光诱导人角膜上皮细胞氧化应激:不同药用植物混合物的乙醇提取物的保护作用。

Blue light-induced oxidative stress in human corneal epithelial cells: protective effects of ethanol extracts of various medicinal plant mixtures.

机构信息

Department of Dermatology, Chonnam National University Medical School and Hospital, Gwangju, Korea.

Department of Chemical Engineering, Sunchon National University, Chonnam, Korea.

出版信息

Invest Ophthalmol Vis Sci. 2014 Jun 12;55(7):4119-27. doi: 10.1167/iovs.13-13441.

Abstract

PURPOSE

To investigate the effects of visible light on human corneal epithelial cells and the impact of natural antioxidants on oxidative stress produced by overexposure to light.

METHODS

Light-emitting diodes with various wavelengths (410-830 nm) were used to irradiate human corneal epithelial cells, and cell viability was assessed. The production of reactive oxygen species (ROS) was analyzed using 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA). Ethyl alcohol (EtOH) extracts were prepared from mixtures of medicinal plants. After application of the EtOH extracts, the free radical scavenging activity was measured using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. The induction of antioxidant enzymes including heme oxygenase-1 (HO-1), peroxiredoxin-1 (Prx-1), catalase (CAT), and superoxide dismutase-2 (SOD-2) by the extracts was evaluated by reverse transcription-polymerase chain reaction and Western blotting. The ability of the extracts to inhibit ROS was also analyzed using DCF-DA.

RESULTS

The viability of corneal epithelial cells was diminished after irradiation of blue light (above 10 J at 410 nm and 50 J at 480 nm). Reactive oxygen species production was induced by irradiation at 410 and 480 nm at doses of 5 J/cm(2) and higher. Ethyl alcohol extracts had potent radical scavenging activity. Application of the extracts not only increased the expression of HO-1, Prx-1, CAT, and SOD-2, but it also attenuated the ROS production induced by blue light in a dose-dependent manner.

CONCLUSIONS

Overexposure to blue light (410-480 nm) may have a harmful effect on human corneal epithelial cells compared with other visible light wavelengths. Medicinal plant extracts can have potent protective effects on blue light-induced oxidative stress.

摘要

目的

研究可见光对人眼角膜上皮细胞的影响,以及天然抗氧化剂对光过度暴露产生的氧化应激的影响。

方法

使用不同波长(410-830nm)的发光二极管照射人眼角膜上皮细胞,评估细胞活力。使用 2',7'-二氯二氢荧光素二乙酸酯(DCF-DA)分析活性氧(ROS)的产生。从药用植物混合物中制备乙醇(EtOH)提取物。应用 EtOH 提取物后,使用 2,2-二苯基-1-苦基肼(DPPH)自由基清除测定法测量自由基清除活性。通过逆转录-聚合酶链反应和 Western 印迹评估提取物对血红素加氧酶-1(HO-1)、过氧化物酶-1(Prx-1)、过氧化氢酶(CAT)和超氧化物歧化酶-2(SOD-2)等抗氧化酶的诱导作用。还使用 DCF-DA 分析了提取物抑制 ROS 的能力。

结果

蓝光(410nm 时超过 10J,480nm 时超过 50J)照射后,角膜上皮细胞活力降低。在 410nm 和 480nm 下,剂量为 5J/cm2 及更高时,会诱导 ROS 产生。乙醇提取物具有很强的自由基清除活性。提取物的应用不仅增加了 HO-1、Prx-1、CAT 和 SOD-2 的表达,而且还以剂量依赖的方式减弱了蓝光诱导的 ROS 产生。

结论

与其他可见光波长相比,过度暴露于蓝光(410-480nm)可能对人眼角膜上皮细胞有害。药用植物提取物对蓝光诱导的氧化应激具有很强的保护作用。

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