茶多酚对UVA/UVB所致HSF细胞急性光损伤的防护作用  

Protective Effect of Tea Polyphenols on Acute Photodamage of Human Skin Fibroblast Cells Induced by UVA/UVB

作　　者：刘清 江娜 彭丽倩[1] 梁碧华 张尔婷 李润祥 邓蕙妍 李华平 李振洁 朱慧兰 

机构地区：[1]广州市皮肤防治所,广州市510095 [2]广州医科大学附属第五医院 [3]广州市第一人民医院

出　　处：《中国激光医学杂志》2017年第5期246-253,290,共9页Chinese Journal of Laser Medicine & Surgery

基　　金：广东省2014年科技计划项目(2014A020212570);2016年广州市科技计划项目(201604020093)

摘　　要：目的探讨茶多酚对长波紫外线(ultraviolet A,UVA)、中波紫外线(ultraviolet B,UVB)所致人皮肤成纤维细胞(human skin fibroblasts,HSF)急性光损伤的防护作用及其对Ⅱ相解毒酶及抗氧化酶mRNA表达的影响。方法不同浓度茶多酚(0、2.5、5、10、20和40 g/ml)孵育HSF细胞24 h,MTT法检测各细胞增殖活性,筛选适宜浓度茶多酚;将HSF细胞分为空白组、药物组、UVA照射组、UVB照射组、UVA+茶多酚组、UVB+茶多酚组,采用2'7'-二氯荧乙酰乙酸盐(2'7'-dichlorofluorescin diacetate,DCFH-DA)荧光探针检测细胞内活性氧簇(reactive oxygen species,ROS)水平,酶生化法测定各组乳酸脱氢酶(lactate dehydrogenase,LDH)释放水平,单细胞凝胶电泳法检测DNA损伤情况,采用实时荧光定量PCR(Quantitative Real-time PCR,qRT-PCR)检测核因子E2相关因子2(Nuclear factor erythroid 2-related factor 2,Nrf2)信号通路下游各Ⅱ相解毒酶mRNA的表达情况。结果茶多酚浓度为5μg/ml时,对HSF细胞增殖活性无明显影响,故后续实验采用5μg/ml浓度。与空白对照组相比,UVA及UVB照射组ROS水平、LDH释放水平明显升高,DNA损伤加重;与单纯光照组相比,加茶多酚预处理后行光照组ROS水平、LDH释放水平降低,DNA损伤减轻,差异具有统计学意义(P<0.05);与空白对照组相比,药物组过氧化氢酶(catalase,CAT)、谷氨酸-半胱氨酸连接酶催化亚基(γ-glutamyl cystine ligase catalytic subunit,GCLC)、谷氨酸-半胱氨酸连接酶调节亚基(γ-glutamyl cystine ligase modulatory subunit,GCLM)、超氧化物歧化酶(superoxide dismutase,SOD)、醌氧化还原酶1(Nuinone oxidoreductase 1,NQO1)的mRNA表达均增加(P<0.05)。结论茶多酚能有效清除ROS,降低LDH水平,减轻DNA损伤,防护UV致HSF细胞急性光损伤,机制可能与其促进Ⅱ相解毒酶及抗氧化物酶mRNA表达相关。Objective To investigate the protective effects of tea polyphenols on the HSF cells suffered the acute light damage of UVA/UVB and their effect on phase Ⅱ and antioxidant enzyme mRNA expression. Methods HSF cells were incubated with tea polyphenols of different concentrations （0, 2. 5, 5, 10, 20 and 40 g/ml） for 24 h, the proliferation of the cell activity was detected with MTT and the suitable concentrations of tea polyphenols were selected. The HSF cells were divided into six groups ： the blank control group, TP group, UVA radiation group, UVB radiation group, UVA ＋ TP group and UVB ＋ TP group. The level of ROS, the level of LDH, the DNA damage, the expression of the mRNA Ⅱ phase detoxifying enzyme was detected with DCFH-DA fluorescent probe method, with biochemical methods, with comet assay and with qRT-PCR respectively. Results The tea polyphenols concentration of 5 μg/ml was adopted in the experiment followed as no significant effect on the proliferation of HSF cells was noticed at this concentration. The ROS levels, LDH levels and DNA injury of the UVA group and UVB group were dramatically higher than those of the control group. The TP groups had a lower ROS level, LDH level and DNA injury than the pure UV groups, and the difference was statistically significant （ P 〈 0.05 ）. The expression of CAT, GCLM, GCLC, SOD and NQO1 mRNA of the drug groups was stronger than that of the blank control group （ P 〈 0. 05 ）. Conclusions TP has protective effect on HSF cells injured due to UV induced oxidative stress, including eliminating ROS and LDH and reducing DNA injury. The mechanism may be related to promote the expression of phase Ⅱ detoxifying enzyme and antioxidant en- zyme mRNA.

关 键 词：茶多酚 紫外线 HSF细胞 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]