摘要
目的探讨紫外线诱导表皮细胞细胞凋亡过程中活性氧的含量及超氧化物歧化酶对活性氧的抑制作用及对凋亡的影响.方法取含10%小牛血清的RMPI1640培养基培养表皮细胞,定量接种于培养皿或96孔板中.将接种于96孔培养板的表皮细胞分为对照组、UV照射组、UV+SOD组.紫外线照射后采用2',7'-二氢二氯荧光黄双乙酸钠(DCFH-DA)为标记探针,通过流式细胞术检测三组细胞内2',7'-二氢二氯荧光黄(DCF)的荧光强度,间接检测细胞内活性氧的水平.以AnnexinV/PI双染色法通过流式细胞仪检测三组表皮细胞凋亡百分率.结果 (1)细胞凋亡率:对照组(13.0205±0.6144)%;UV照射组(34.0426±1.2427)%;UV+SOD组(11.5324±0.6248)%;(2)细胞活性氧水平:对照组(85.0231±1.2134)%;UV照射组(96.2402±0.5402)%;UV+SOD组(89.3625±0.5402)%.结论紫外线能升高表皮细胞的SOD,可减轻紫外线诱导ROS产生,作用细胞凋亡抑制ROS水平升高.
Objective To study the content of reactive oxygen species in（ROS） in the apoptosis of epidermal cells induced by ultraviolet（UV）,the inhibiory effects of superoxide dismut-ase（SOD） on ROS,and the effects of SOD on apoptosis.Methods Epidermal cells were cultured in RMPI1640 medium supplemented with 10% calf serum,and then were inoculated in petri dishes or 96-well plates.Epidermal cells in 96-well plates were randomly divided into three groups：control group,UV irradiation group and UV ＋ SOD group.UV was irradiated in cultural medium with or without SOD except control group.Epidermal cells were harvested to esti mate the intracellular ROS levels by detecting the fluorescent intensity of the 2＇,7＇-dichlorofluorescein diacetate（DCFH-DA） by flowcytometry.Annexin V/PI two-color flowcytometry was used to detect the apoptosis of epidermal cells.Results（1） The apoptotic rates of cells in the control group,UV irradiation group,and UV＋SOD group were（13.020 5±0.614 4）%,（34.042 6±1.242 7）%,and（11.532 4±0.624 8）%,respectively.There was a significant difference between UV irradiation group and the other two groups（P 0.05）.（2） The levels of reactive oxygen species in the control group,UV irradiation group,and UV＋SOD group were（85.023 1±1.213 4）%,（96.240 2±0.540 2）%,and（89.362 5±0.540 2）%,respectively.There was a significant difference between UV irradiation group and the other two groups（P〈0.05）.Conclusion Ultraviolet irradiation can injure epidermal cells.This study confirm that antioxidants can reduce the initial UV-induced ROS generation and inhibit the apoptosis of cells,and provide an indiredtive proof for the positive correlation be-tween ROS and apoptosis.
出处
《昆明医学院学报》
2010年第4期36-39,共4页
Journal of Kunming Medical College
关键词
紫外线
活性氧
表皮细胞
凋亡
Ultraviolet
Reactive oxygen species
Epidermal cells
Apoptosis
作者简介
徐自祥（1972～），男，云南陆良县人，医学学士，主治医师，主要从事口腔科临床工作．
