Protective effect of resveratrol on UVA-irradiated HaCaT cells and its mechanism

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Protective effect of resveratrol on UVA-irradiated HaCaT cells and its mechanism

RADIOBIOLOGY AND RADIOMEDICINE | 更新时间：2021-12-10

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- Protective effect of resveratrol on UVA-irradiated HaCaT cells and its mechanism
- Journal of Radiation Research and Radiation Processing Vol. 35, Issue 2, Pages: 20201(2017)
- 作者机构：
  
  1.中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中国疾病预防控制中心重点实验室北京 100088
- 作者简介：
- 基金信息：
- DOI：10.11889/j.1000-3436.2017.rrj.35.020201
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Chen LI, Chunyan WANG, Mei TIAN, et al. Protective effect of resveratrol on UVA-irradiated HaCaT cells and its mechanism. [J]. Journal of Radiation Research and Radiation Processing 35(2):20201(2017)
DOI：

Chen LI, Chunyan WANG, Mei TIAN, et al. Protective effect of resveratrol on UVA-irradiated HaCaT cells and its mechanism. [J]. Journal of Radiation Research and Radiation Processing 35(2):20201(2017) DOI： 10.11889/j.1000-3436.2017.rrj.35.020201.

摘要

采用Cell Counting Kit-8（CCK-8试剂盒）检测HaCaT细胞的增殖水平。Annexin V-FITC/PI流式细胞术检测HaCaT细胞的凋亡率和死亡率。单丹磺酰尸胺（Monodansylcadaverine, MDC）荧光染色和二氯荧光黄二乙酸酯（Dichlorodihydrofluorescein diacetate, DCFH-DA）探针标记后，用激光共聚焦显微镜进行扫描，分别检测HaCaT细胞内自噬体和活性氧（Reactive oxygen species, ROS）水平变化。Western blot检测HaCaT细胞内LC3B-II、磷酸化AKT（p-AKT）和总AKT蛋白表达变化。结果显示：经10~80 J/cm,2, UVA照射后，HaCaT细胞的增殖水平随受照剂量的增加逐渐降低（,p,<, 0.05）；白藜芦醇预处理能显著促进UVA照射后HaCaT细胞增殖（,p,<, 0.01），降低其凋亡率和死亡率（,p,<, 0.05）；经20 J/cm,2, UVA照射后3~24 h，HaCaT细胞内LC3B-II蛋白表达增强（,p,<, 0.01）；白藜芦醇预处理能使UVA照射后HaCaT细胞内自噬增强（,p,<, 0.01），ROS水平降低（,p,<, 0.01），p-AKT蛋白表达增强（,p,<, 0.01）。提示白藜芦醇可能通过促进细胞自噬和AKT蛋白磷酸化，以及降低细胞内ROS水平，拮抗UVA导致的HaCaT细胞增殖抑制、凋亡和死亡。

Abstract

The Cell Counting Kit-8 (CCK-8) was used to detect the cell survival rate of HaCaT cells. The apoptosis rate and mortality of HaCaT cells were detected using flow cytometry with the Annexin VFITC and PI double labeling method. Autophagosomes and the generation of reactive oxygen species (ROS) in HaCaT cells were monitored by laser scanning confocal microscopy using monodansylcadaverine (MDC) and dichlorodihydrofluorescein diacetate (DCFH-DA) staining, respectively. The expression of LC3B-II, p-AKT, and total AKT protein in HaCaT cells was determined by western blotting. The results showed that HaCaT cell proliferation was inhibited gradually (,p,<, 0.05) after irradiation with a dose of 10~80 J/cm,2, UVA. Treatment with resveratrol enhanced the proliferation of UVA-irradiated HaCaT cells (,p,<, 0.01) and decreased their apoptosis rate and mortality (,p,<, 0.05). The expression of LC3B-II protein was increased (,p,<, 0.01) after irradiation with 20 J/cm,2, UVA for 3~24 h. After treatment with resveratrol, autophagy was enhanced (,p,<, 0.01), intracellular ROS level was decreased (,p,<, 0.01), and p-AKT expression was significantly increased (,p,<, 0.01) in UVA-irradiated HaCaT cells. These results indicate that resveratrol could reduce UVA-induced proliferative inhibition, apoptosis, and death in HaCaT cells by enhancing the autophagic process and p-AKT protein expression and by reducing ROS production.

关键词

白藜芦醇HaCaT细胞UVA防护效应

Keywords

ResveratrolHaCaT cellsUVAProtective effect

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Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Peking Union Medical College

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（北京协和医学院中国医学科学院放射医学研究所）

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