UVB辐射HaCaT细胞损伤模型的建立  

Model establishment of UVB-induced damage in Ha Ca T cell

作　　者：高薇[1] 侯微[1] 李伟[1,2] 王玉帅[1] 刘清秀[1] 王英平[1] 

机构地区：[1]中国农业科学院特产研究所新药研究室,长春130112 [2]吉林农业大学中药材学院,长春130118

出　　处：《中药药理与临床》2014年第5期136-139,共4页Pharmacology and Clinics of Chinese Materia Medica

摘　　要：目的:UVB辐射细胞损伤程度依赖于辐射剂量及辐射后孵育的时间,观察不同辐射剂量及辐射后孵育不同时间的细胞损伤程度,可以建立适宜的UVB损伤模型,为天然抗紫外药物的筛选奠定基础,有助于天然紫外防护品的开发。方法:分别用10mj/cm2、20 mj/cm2、30 mj/cm2、40 mj/cm2的剂量辐射细胞,辐射后孵育不同时间,采用MTT法检测细胞活性、流式细胞术检测细胞调亡率以及彗星电泳试验观察细胞DNA损伤程度;对于优化的模型条件用阳性药维生素C进一步验证。结果:发现用30 mj/cm2以上的剂量辐射细胞,辐射后孵育24 h可明显观察到DNA损伤,且该剂量下细胞凋亡率显著升高,维生素C对该条件建立的模型有明显的保护作用。结论:采用30 mj/cm2的辐射剂量,辐射后孵育24h条件建立模型。Objective： The extent of cell damage relies on the radiation dose and incubation time after radiation. The observation of the damage degrees of the cells after different radiation doses and different incubation time,we can establish suitable UVB damage model to screen natural uvioresistant drugs and develop natural UV protection products. Methods： The Ha Ca T cells were radiated by different doses of UVB： 10 mj /cm2,20mj/cm2,30mj/cm2,40mj/cm2,then incubated by different time. MTT was used to detect the cell viability,flow cytometry was used to assay the cell apoptosis rate and the cellular DNA damage was observed by comet electrophoresis. The optimized model conditions were further verified by positive medicine-Vc. Results： We found that the cells which were incubated by 24 h after exceeding 30 mj / cm2 radiation could generate obvious DNA damage,meanwhile,under which condition the cell apoptosis rate was increased significantly. Positive medicine（ Vc）has obvious protective effect. Conclusions： The cells which were incubated by 24 h after the radiation dose of 30 mj / cm2 is an ideal UV damage model.

关 键 词：UVB HA Ca T细胞 彗星电泳实验 流式细胞术 

分 类 号：R285[医药卫生—中药学]