摘要
通过常规方法培养可稳定表达绿色荧光蛋白(GFP)的HK-2细胞,应用Western blotting和荧光显微镜分析肾毒性药物阿米卡星对HK-2细胞GFP表达及荧光强度的影响。通过测算已知的4种中药肾毒性成分(汉防已碱、马兜铃酸、雷公藤甲素、芦荟大黄素)的IC50和FC50,分析两者的相关性。结果显示阿米卡星可诱发HK-2细胞内的荧光产生并在24 h内逐渐增强,GFP蛋白的表达在24 h时明显增加(与对照组比,P<0.01)。4种中药肾毒性成分的IC50和FC50之间具有良好的相关性。稳定表达GFP的HK-2细胞内的荧光强度的变化可以较好地反映细胞损伤程度。该方法的建立为中药成分的肾毒性的检测和早期预测提供了有力的工具。
A method was established to detect the nephrotoxicity of traditional Chinese medicine ingredients.HK-2 cells with stable expression of green fluorescent protein（GFP） were cultured routinely.Western blotting and fluorescence microscopy analysis were selected to analyze the effects of amikacin with nephrotoxicity on expression of GFP and intracellular fluorescence intensity in HK-2 cells.The values of IC50 and FC50 of four known traditional Chinese medicine（triptolide,aristolochic acid,tetrandrine,aloe-emodin） ingredients with nephrotoxicity were estimated respectively;and the correlation between IC50 and FC50 were analyzed.The results showed that the fluorescence in HK-2 cells induced by amikacin was gradually increased within 24 h;GFP expression was significantly increased at 24 h（compared with the control group,P0.01）.The values of FC50 obtained from four known traditional Chinese medicine ingredients with nephrotoxicity were in positive correlation with the corresponding values of IC50.It can be concluded that the changes of fluorescence intensity in HK-2 cells with stable expression of GFP can reflect the degree of cell damage.This method will provide potential alternatives for the detection and early prediction of nephrotoxicity of traditional Chinese medicine ingredients.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2012年第6期526-529,共4页
Journal of China Pharmaceutical University
基金
江苏省中药药效与安全性评价重点实验室开放课题资助项目(No.T09006)~~
关键词
中药肾毒性
HK-2细胞
热休克蛋白70
绿色荧光蛋白
nephrotoxicity of traditional Chinese medicine
HK-2 cell
heat shock protein 70
green fluorescentprotein
