白藜芦醇对UVA照射HaCaT细胞的防护效应及其机制
Protective effect of resveratrol on UVA-irradiated HaCaT cells and its mechanism
- 辐射研究与辐射工艺学报 2017年35卷第2期 页码：20201
- 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：
[ "李辰, 女, 1987年4月出生, 2012年于首都医科大学获得硕士学位, 药理学专业, 现在中国疾控中心辐射安全所, 助理研究员, 从事辐射防护药物研究, E-mail:lichen317@163.com", "First author: LI Chen (female) was born in April 1987 and received her master's degree from Capital Medical University, majoring in pharmacology. Now she works in NIRP as an assistant professor, engaging in radiation protection drugs, E-mail:lichen317@163.com" ]
苟巧, 博士, 副研究员, E-mail:xiaogou824@163.comPh.D. GOU Qiao, associate professor, E-mail:xiaogou824@163.com
- 基金信息：
DOI：10.11889/j.1000-3436.2017.rrj.35.020201
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李辰, 王春燕, 田梅, 等. 白藜芦醇对UVA照射HaCaT细胞的防护效应及其机制[J]. 辐射研究与辐射工艺学报, 2017,35(2):20201.
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, 2017,35(2):20201.
李辰, 王春燕, 田梅, 等. 白藜芦醇对UVA照射HaCaT细胞的防护效应及其机制[J]. 辐射研究与辐射工艺学报, 2017,35(2):20201. DOI： 10.11889/j.1000-3436.2017.rrj.35.020201.
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, 2017,35(2):20201. 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
references
S Hu , F Chen , M Wang . Photoprotective effects of oxyresveratrol and Kuwanon O on DNA damage induced by UVA in human epidermal keratinocytes . Chemical Research Toxicology , 2015 . 28 (3 ):541 -548 . DOI:10.1021/tx500497uhttp://doi.org/10.1021/tx500497u .
M Grigalavicius , J Moan , A Dahlback , 等 . Daily, seasonal, and latitudinal variations in solar ultraviolet A and B radiation in relation to vitamin D production and risk for skin cancer . International Journal of Dermatology , 2016 . 55 (1 ):e23 -28 . DOI:10.1111/ijd.13065http://doi.org/10.1111/ijd.13065 .
K Hiramoto , Y Yamate . Gp91phox-derived reactive oxygen species/urocortin 2/corticotropin-releasing hormone receptor type 2 play an important role in long-term ultraviolet a eye irradiation-induced photoaging . Journal of Photochemistry and Photobiology , 2016 . 92 (1 ):180 -186 . DOI:10.1111/php.12553http://doi.org/10.1111/php.12553 .
Y Feng , J Zhou , Y Jiang . Resveratrol in lung cancer-a systematic review . Journal of Balkan Union of Oncology , 2016 . 21 (4 ):950 -953 . http://www.pubfacts.com/detail/27685918/Resveratrol-in-lung-cancer-a-systematic-reviewhttp://www.pubfacts.com/detail/27685918/Resveratrol-in-lung-cancer-a-systematic-review, .
S Samarghandian , M Azimi-Nezhad , R Afshari , 等 . Effects of buprenorphine on balance of oxidant/antioxidant system in the different ages of male rat liver . Journal of Biochemical and Molecular Toxicology , 2015 . 29 (6 ):249 -253 . DOI:10.1002/jbt.21691http://doi.org/10.1002/jbt.21691 .
S R Gundala , C H Yang , R Mukkavilli , 等 . Hydroxychavicol, a betel leaf component, inhibits prostate cancer through ROS-driven DNA damage and apoptosis . Toxicology and Applied Pharmacology , 2014 . 280 (1 ):86 -96 . DOI:10.1016/j.taap.2014.07.012http://doi.org/10.1016/j.taap.2014.07.012 .
孙 颖 , 黄 明亮 , 王 华 . 白藜芦醇抗氧化性及安全性的研究进展 . 食品工业 , 2013 . 34 (4 ):177 -180 . http://www.cnki.com.cn/Article/CJFDTOTAL-SPGY201304066.htmhttp://www.cnki.com.cn/Article/CJFDTOTAL-SPGY201304066.htm, .
Ying SUN , Mingliang HUANG , Hua WANG . Review on antioxidant activity and safety of resveratrol . The Food Industry , 2013 . 34 (4 ):177 -180 . http://www.cnki.com.cn/Article/CJFDTOTAL-SPGY201304066.htmhttp://www.cnki.com.cn/Article/CJFDTOTAL-SPGY201304066.htm, .
J Moscat , M Karin , M T Diaz-Meco . p62 in Cancer: signaling adaptor beyond autophagy . Cell , 2016 . 167 (3 ):606 -609 . DOI:10.1016/j.cell.2016.09.030http://doi.org/10.1016/j.cell.2016.09.030 .
G Meyer , A Czompa , C Reboul , 等 . The cellular autophagy markers Beclin-1 and LC3B-II are increased during reperfusion in fibrillated mouse hearts . Current Pharmaceutical Design , 2013 . 19 (39 ):6912 -6918 . DOI:10.2174/138161281939131127122510http://doi.org/10.2174/138161281939131127122510 .
L Tang , F Yang , Z Fang , 等 . Resveratrol ameliorates alcoholic fatty liver by inducing autophagy . American Journal of Chinese Medicine , 2016 . 44 (6 ):1207 -1220 . DOI:10.1142/S0192415X16500671http://doi.org/10.1142/S0192415X16500671 .
H Deng , M T Mi . Resveratrol attenuates Aβ25-35 caused neurotoxicity by inducing autophagy through the TyrRS-PARP1-SIRT1 signaling pathway . Neurochemical Research , 2016 . 41 (9 ):2367 -2379 . DOI:10.1007/s11064-016-1950-9http://doi.org/10.1007/s11064-016-1950-9 .
G A Abdel-Aleem , E F Khaleel , D G Mostafa , 等 . Neuroprotective effect of resveratrol against brain ischemia reperfusion injury in rats entails reduction of DJ-1 protein expression and activation of PI3K/Akt/GSK3b survival pathway . Archives of Physiology Biochemistry , 2016 . 122 (4 ):200 -213 . DOI:10.1080/13813455.2016.1182190http://doi.org/10.1080/13813455.2016.1182190 .
Z X Dai , P F Lei , J Xie , 等 . Antitumor effect of resveratrol on chondrosarcoma cells via phosphoinositide 3-kinase/AKT and p38 mitogen-activated protein kinase pathways . Molecular Medicine Reports , 2015 . 12 (2 ):3151 -3155 . DOI:10.3892/mmr.2015.3683http://doi.org/10.3892/mmr.2015.3683 .
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