In vitro study of the nephrotoxicity of total Dahuang(Radix Et Rhizoma Rhei Palmati) anthraquinones and emodin in monolayer human proximal tubular epithelial cells cultured in a transwell chamber  

作　　者：Cao Chunyu Hui Lianqiang Li Chun Yang Yifei Zhang Jiyuan Liu Ting Hao Ran Zhang Yi 

机构地区：[1]Safety Evaluation Center for Chinese Materia Medica,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China [2]Quality Standard Center for Chinese Materia Medica, Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China [3]Department of Sciences and Education,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China [4]Pharmacology Research Center for Chinese Materia Medica,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China

出　　处：《Journal of Traditional Chinese Medicine》2019年第5期609-623,共15页中医杂志（英文版）

基　　金：Supported by the Central Public Welfare Research Institutes Foundation-funded Project:A new research of renal toxicity early detection of Chinese medicine(ZZ0908035);Establishment of HK-2 cell monolayer model of human proximal renal tubular epithelial cells(ZXKT15022);Bioactivity based quality control for Chinese herbal medicine using Rhubarb as model system(GH2017-01-02);Natural Science Foundation-funded Project:Study on the bacterial endotoxin detection method of traditional Chinese medicine injections(90709043)

摘　　要：OBJECTIVE: To examine changes in the morphology and physiological functions of human proximal tubular epithelial cells (HK-2 cells) caused by total Dahuang (Radix Et Rhizoma Rhei Palmati) anthraquinones (TDA) and emodin. METHODS: HK-2 cells were cultured on polycarbonate (PCF) membranes to form a complete monolayer of cells. A fluorescein isothiocyanate- dextran (FITC) permeability assay was conducted and secretion of γ-glutamyltranspeptidase (GGT), lactate dehydrogenase (LDH), N-acetyl-β-D-glucosaminidase (NAG) and kidney injury molecule 1 (KIM-1) was examined. The reabsorption of glucose and the excretion of para-aminohippuric acid (PAH) by HK-2 cells were also examined. The morphology of HK-2 cells was observed using optical microscopy and scanning electron microscopy. The cytoskeleton of HK-2 cells was observed under a fluorescence microscope. RESULTS: Compared with the results for the dimethyl sulfoxide group, treatment of cells with TDA and emodin showed statistically significant differences in the FITC leakage rate, the apical / basolateral ratio of LDH and GGT, and the secretion of GGT, LDH, NAG and KIM-1. At 64 μg/mL, TDA markedly inhibited blood glucose reabsorption and remarkably suppressed PAH excretion by HK-2 cells. Both TDA and emodin caused various degrees of damage to the morphology and cytoskeleton of HK-2 cells with the degree of damage correlating positively with the dosage of the tested substances.CONCLUSION: Both TDA and emodin caused damage to human renal proximal tubular epithelial cells at certain dosages. At the same dosage, TDA caused more severe damage than emodin to the HK-2 cells.OBJECTIVE: To examine changes in the morphology and physiological functions of human proximal tubular epithelial cells(HK-2 cells) caused by total Dahuang(Radix Et Rhizoma Rhei Palmati) anthraquinones(TDA) and emodin.METHODS: HK-2 cells were cultured on polycarbonate(PCF) membranes to form a complete monolayer of cells.A fluorescein isothiocyanatedextran(FITC) permeability assay was conducted and secretion of γ-glutamyltranspeptidase(GGT),lactate dehydrogenase(LDH), N-acetyl-β-D-glucosaminidase(NAG) and kidney injury molecule 1(KIM-1) was examined.The reabsorption of glucose and the excretion of para-aminohippuric acid(PAH)by HK-2 cells were also examined.The morphology of HK-2 cells was observed using optical microscopy and scanning electron microscopy.The cytoskeleton of HK-2 cells was observed under a fluorescence microscope.RESULTS: Compared with the results for the dimethyl sulfoxide group, treatment of cells with TDA and emodin showed statistically significant differences in the FITC leakage rate, the apical/basolateral ratio of LDH and GGT, and the secretion of GGT,LDH, NAG and KIM-1.At 64 μg/mL, TDA markedly inhibited blood glucose reabsorption and remarkably suppressed PAH excretion by HK-2 cells.Both TDA and emodin caused various degrees of damage to the morphology and cytoskeleton of HK-2 cells with the degree of damage correlating positively with the dosage of the tested substances.CONCLUSION: Both TDA and emodin caused damage to human renal proximal tubular epithelial cells at certain dosages.At the same dosage, TDA caused more severe damage than emodin to the HK-2 cells.

关 键 词：ANTHRAQUINONES RHEUM EMODIN NEPHROTOXICITY Human proximal tubular epithelial cell TRANSWELL CHAMBER 

分 类 号：R[医药卫生]