Method for in vitro detection of nephrotoxicity of traditional Chinese medicine ingredients

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,P<0.01).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.