紫檀芪对长波紫外线辐射所致人成纤维细胞急性光损伤的保护作用  

Protective Effects of Pterostilbene on Ultraviolet A-induced Photo-damage in Human Foreskin Fibroblasts

作　　者：陈荃 李华平 张尔婷 梁碧华 邓蕙妍 李润祥 彭丽倩[2] 莫子茵 代歆悦 朱慧兰 CHEN Quan1 , LI Huaping1, ZHANG Erting2, LIANG Bihua1 , DENG Huiyan1 , LI Runxiang1 , PENG Liqian2, MO Ziyin2, DAI Xinyue2, ZHU Huilan1(1. Guangzhou Institute of Dermatology, Guangzhou 510095, China; 2. Guangzhou Medical Univer- sity, Guangzhou 510182, China)

机构地区：[1]广州市皮肤病防治所,广州医科大学皮肤病研究所,广东广州510095 [2]广州医科大学,广东广州510182

出　　处：《中国皮肤性病学杂志》2018年第4期375-381,共7页The Chinese Journal of Dermatovenereology

基　　金：广东省科技计划项目(2014A020212570,2014A020212649);广州市科技计划项目(201604020093,201607010382);广州市卫生局医药卫生科技项目(20171A011282,20171A011287)

摘　　要：目的探讨紫檀芪(pterostilbene)对长波紫外线(UVA)辐射所致的人成纤维细胞急性损伤的保护作用,初步分析其分子机制。方法 MTT法测定紫檀芪对人皮肤成纤维细胞活力的影响,紫檀芪与人皮肤成纤维细胞共培养24h后,以20J/cm2的UVA剂量进行照射,然后采用乳酸脱氢酶释放法测定细胞活力,DCFH-DA测定胞内活性氧(cellular reactive oxygen species,ROS)水平,彗星实验检测DNA损伤,Western印迹检测核因子NF-E2相关因子2(nuclear factor erythroid 2-related factor2,Nrf2)蛋白细胞定位以及siRNA沉默Nrf2。结果与对照相比,UVA照射后乳酸脱氢酶相对释放率增加为164.53%(P<0.001)。而UVA照射前4μmol/L和8μmol/L紫檀芪预处理细胞24h,则细胞乳酸脱氢酶相对释放率分别恢复到对照组的100.11%(P>0.05)和98.69%(P>0.05)。流式细胞术分析显示,经UVA辐照后,其ROS水平上升至对照细胞的264.4%(P<0.001),UVA照射前4μmol/L紫檀芪预处理细胞24h,则活性氧(ROS)水平恢复至对照细胞的113.50%(P<0.05)。彗星实验结果显示,空白对照组细胞的尾部DNA百分比为(1.795±2.147)%,UVA辐照组为(65.50±12.418)%,与空白对照组相比DNA损伤显著增加(t=40.68,P<0.000 1);而紫檀芪预处理后再接受UVA辐照组为(13.53±8.152)%,与UVA辐照组相比其DNA损伤显著减轻(t=27.85,P<0.000 1)。紫檀芪处理提高了细胞核中的Nrf2蛋白水平。转染siRNA-Nrf2后,细胞活性与对照细胞差异无统计学意义(P>0.05),但经UVA照射,紫檀芪的预处理细胞活性为对照组的35.32%(P<0.000 1),紫檀芪的保护作用显著降低。结论紫檀芪能有效防御UVA所致成纤维细胞急性光损伤。这种保护作用可能与紫檀芪激活Nrf2信号通路相关。Objective To evaluate the photoprotective effects of pterostilbene against UVA induced acute photo-damage in human foreskin fibroblasts（HFF）, and to investigate its underlying molecular mechanism.Methods HFF cells were pretreated with pterostilbene before being irradiated with 20J/cm2 ultraviolet A（UVA）and then incubated for 24h.Then cell viability was assessed by lactate dehydrogenase release assay, DCFH-DA assay was performed to evaluate cellular reactive oxygen species（ROS）generation, comet assay to observe DNA damage, Western blot to analyze the cellular location of NF-E2-related factor 2（Nrf2）.Additionally, the Nrf2 knockdown was performed by small interference RNA（siRNA）transfection.Results The lactate dehydrogenase release increased to 164.53%（P〈0.001）of control cell after UVA irradiation, whereas 4μmol/L and 8μmol/L pretreatment with pterostilbene decreased to 100.11%（P〉0.05）and 98.69%（P〉0.05）respectively.UVA irradiation also induced ROS to 264.4%（P〈0.001）of control, however, pretreatment with pterostilbene decreased the ROS production to 113.5%（P〈0.05）of control.Quantification of comet tails indicated that control cells were 1.80±2.15, UVA-irradiation cells were 65.50±12.42（P〈0.000 1）, pterostilbene pretreated cells were 13.53±8.15（P〈0.000 1）.Pretreatment with pterostilbene also increase the nuclear protein level of Nrf2.Transfection with Nrf2 targeted siRNA alone without UVA irradiation had no obvious effect on the cell viability（P〉0.05）, while the viability of Nrf2 silencing cells pretreated with pterostilbene prior to UVA irradiation decreased to 35.32%（P〈0.000 1）of control.Conclusion Pterostilbene could protect against UVA-induced photo-damage, partially through activation of Nrf2 pathway in HFF cells.

关 键 词：紫檀芪 长波紫外线 人皮肤成纤维细胞 氧化损伤 NRF2 

分 类 号：R751[医药卫生—皮肤病学与性病学]