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白藜芦醇可预防活性氧诱导的发光二极管产生的蓝光对人皮肤成纤维细胞的影响。

Resveratrol Prevents Reactive Oxygen Species-Induced Effects of Light-Emitting Diode-Generated Blue Light in Human Skin Fibroblasts.

作者信息

Mamalis Andrew, Koo Eugene, Jagdeo Jared

机构信息

*Department of Dermatology, University of California at Davis, Sacramento, California; †Dermatology Service, Sacramento VA Medical Center, Mather, California; ‡Department of Dermatology, SUNY Downstate Medical Center, Brooklyn, New York.

出版信息

Dermatol Surg. 2016 Jun;42(6):727-32. doi: 10.1097/DSS.0000000000000744.

Abstract

BACKGROUND

Light-emitting diode-generated blue light (LED-BL) is part of the visible light spectrum that does not cause DNA damage and may represent a safer alternative to ultraviolet phototherapy. Previous research demonstrated that LED-BL can inhibit adult human skin fibroblast proliferation and migration speed and is associated with increased reactive oxygen species (ROS) generation in a dose-dependent manner. In addition, resveratrol possesses potent intracellular antioxidative effects on ROS-free radicals in human skin fibroblasts.

OBJECTIVE

The authors studied the effects on migration speed as a surrogate to measure LED-BL effects on fibroblast function. The authors hypothesized that resveratrol, a potent scavenger of ROS, could prevent the effects of LED-BL on fibroblast migration speed. This would implicate ROS as the mechanistic driver of LED-BL effects on human skin fibroblasts.

METHODS

To demonstrate that resveratrol could prevent the effects of LED-BL (415-nm), fibroblasts were incubated with resveratrol (Sigma-Aldrich, St. Louis, MO) at concentrations of 0.001% and 0.0001% for 24 hours and then irradiated with LED-BL at fluences of 30, 45, and 80 J/cm. Postirradiation fibroblast migratory speed was assayed in an environment-controlled computer-assisted video microscopy system. Reactive oxygen species levels were measured by flow cytometric analysis of dihydrorhodamine. Statistical analyses with analysis of variance and Student t-test were performed to compare individual treatment arms and matched controls.

RESULTS

The experimental results demonstrate that pretreatment of skin fibroblasts with resveratrol at concentrations of 0.001% and 0.0001% prevents the effects of 30, 45, and 80 J/cm of LED-BL on fibroblast migration speed. The authors found that LED-BL at a fluences of 30, 45, and 80 J/cm significantly increased ROS, whereas pretreatment with 0.001% resveratrol significantly reduced ROS generation.

CONCLUSION

The findings demonstrate that LED-BL-induced decreases in fibroblast migration speed can be prevented by pretreating cells with resveratrol. This finding supports the hypothesis that ROS generation is the most likely driver of LED-BL-induced alterations in migration speed and suggests that ROS generation may be responsible for a number of other alterations seen after LED-BL phototherapy, such as decreases in cellular migration, cytokine levels, and myofibroblast differentiation. The authors hypothesize that their findings may result in greater understanding of the fundamental mechanisms underlying visible light interaction with skin and they hope dermatologists and other researchers may use these pathways for patient benefit.

摘要

背景

发光二极管产生的蓝光(LED-BL)是可见光光谱的一部分,不会造成DNA损伤,可能是紫外线光疗更安全的替代方法。先前的研究表明,LED-BL可抑制成人人类皮肤成纤维细胞的增殖和迁移速度,并以剂量依赖的方式增加活性氧(ROS)的生成。此外,白藜芦醇对人类皮肤成纤维细胞中的ROS自由基具有强大的细胞内抗氧化作用。

目的

作者研究了对迁移速度的影响,以此作为衡量LED-BL对成纤维细胞功能影响的替代指标。作者假设,作为ROS的有效清除剂,白藜芦醇可以预防LED-BL对成纤维细胞迁移速度的影响。这将表明ROS是LED-BL对人类皮肤成纤维细胞产生影响的机制驱动因素。

方法

为了证明白藜芦醇可以预防LED-BL(415纳米)的影响,将成纤维细胞与浓度为0.001%和0.0001%的白藜芦醇(西格玛奥德里奇公司,密苏里州圣路易斯)孵育24小时,然后用30、45和80 J/cm的LED-BL进行照射。在环境控制的计算机辅助视频显微镜系统中测定照射后成纤维细胞的迁移速度。通过对二氢罗丹明的流式细胞术分析来测量活性氧水平。采用方差分析和学生t检验进行统计分析,以比较各个治疗组和匹配的对照组。

结果

实验结果表明,用浓度为0.001%和0.0001%的白藜芦醇预处理皮肤成纤维细胞可预防30、45和80 J/cm的LED-BL对成纤维细胞迁移速度的影响。作者发现,30、45和80 J/cm的LED-BL显著增加了ROS,而用0.001%白藜芦醇预处理可显著减少ROS的生成。

结论

研究结果表明,用白藜芦醇预处理细胞可以预防LED-BL诱导的成纤维细胞迁移速度降低。这一发现支持了以下假设,即ROS生成是LED-BL诱导迁移速度改变的最可能驱动因素,并表明ROS生成可能是LED-BL光疗后出现的许多其他改变的原因,如细胞迁移、细胞因子水平和肌成纤维细胞分化的降低。作者假设,他们的发现可能会使人们对可见光与皮肤相互作用的基本机制有更深入的了解,他们希望皮肤科医生和其他研究人员可以利用这些途径造福患者。

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