原花青素对紫外线辐射后人表皮黑素细胞光保护机制的研究  

Photo-protection of human melanocytes following UV radiation by Proanthocyanidin-rich extract from Grape seeds in vitro

作　　者：马慧军[1] 訾绍霞[2] 李铀[1] 刘雯[1] 王毅侠[1] 赵广[1] 

机构地区：[1]中国人民解放军皮肤病研究所空军总医院皮肤科,北京100036 [2]首都医科大学宣武医院皮肤性病科,北京100053

出　　处：《中国美容医学》2008年第9期1320-1324,共5页Chinese Journal of Aesthetic Medicine

基　　金：国家自然科学基金(编号30600538);博士后科学基金(20070420574)

摘　　要：目的:研究葡萄籽提取物原花青素(OPC)对紫外线辐射后人表皮黑素细胞内活性氧自由基(ROS)清除、细胞周期变化以及黑素合成关键酶表达的影响,以探讨OPC对黑素细胞的光保护机制。方法:培养的正常表皮黑素细胞经15mJ/cm2紫外线辐射后分别以高、中、低(10、50、100μg/ml)三种浓度的OPC作用24h后,以四甲基偶氮唑蓝(MTT)比色法测定细胞存活率;采用二氯荧光素二酯(DCFH-DA)标记法测定细胞内ROS水平;碘化丙啶(PI)、溴脱氧尿苷(BrdU)双标记结合流式细胞仪检测细胞周期;免疫印迹法测定细胞酪氨酸酶(TYR)、酪氨酸酶相关蛋白1(TRP1)、酪氨酸酶相关蛋白2(TRP2)蛋白含量。结果:OPC可呈浓度依赖性地保护受紫外线辐射的黑素细胞存活率、清除紫外线诱导的黑素细胞内活性氧自由基的产生;≥50μg/mlOPC可减少黑素细胞内TYR、TRP1的蛋白表达量、修复紫外线辐射后下降的TRP2蛋白表达;减少紫外线辐射后G1期细胞比例,增加S期细胞比例。结论:天然寡聚体OPC可能通过抑制细胞内ROS的产生、减少黑素合成关键酶的表达、修复停滞于细胞分裂前期状态的细胞来发挥对黑素细胞的光保护作用。Objective To investigate the mechanism of o melanocytes, we evaluated the effects of cellular cycle, igomers Proanthocyanidins（OPC）on photo-protection of human ntracellular reactive oxidative species （ROS） level and protein level of melanogenic enzymes in cultured human melanocytes following UV-irradiation by OPCs. Methods After treatment with three different dose of OPC, normal human melanocytes were irradiated by 15ml/cm^2 UV light. Then, the viabilities of metanocytes were mesured by MTT assay, the protein analysis of Tyrosinase, tyrosinase related protein 1 （TRP1）,and tyrosinase related protein 2 （TRP2） were observed by western blot. Levels of UV-induced ROS in melanocytes and the responses of cell cycle were also examined by immunofluorescence technique. Results The study has demonstrated that OPC, significantly inhibited the cell dead induced by UV irradiation in a dose-dependent manner and the UV-induced intracellular ROS enhancement was also prevented by addition of OPCs in a dosedependent manner. Meanwhile, OPCs also inhibited the extent of G1 arrest that was induced by UV exposure. A concentration of ≥50μg/ml OPC can decrease the protein level of tyrosinase, TRP1 and TRP2 in UV-irradiated human melanocytes. Conclusions OPC has potential effects of photo-protection on human melanocytes by improving cell viability, scavenging intracellular ROS, adjusting cell cycle and inhibiting protein expression of melanogenic enzymes.

关 键 词：光保护 原花青素 黑素细胞 活性氧自由基 

分 类 号：O813[理学]