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《Chinese Journal of Pharmacology and Toxicology》 2013-01
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Cytotoxic effect of dictamnine on HepG2 cells and its possible mechanism

GUO Xiao-pei1,LI Ai-fang1,ZHANG Bao-xu1,2,WANG Qi1,2(1.Department of Toxicology,School of Public Health,Peking University,Beijing 100191,China; 2.Key Laboratory of State Administration of TCM for Compatibility Toxicology, Beijing 100191,China)  
OBJECTIVE To study the dictamnine-induced hepatotoxicity in vitro and to explore its mechanism.METHODS HepG2 cells were exposed to dictamnine 2.5-800 μmol·L-1 for 24 h,and cell viability was examined by MTT assay.Cell membrane injury was examined by detecting the release rate of lactate dehydrogenase(LDH),and the morphological changes were observed under a contrast microscope.The activities of alanine transaminase(ALT),aspartate aminotransferase(AST),glutathione S-transferase(GST) and glutamyltranspeptidase(GGT) in HepG2 cell cultures were measured using enzyme-labeled instrument,respectively.The mitochondrial membrane potential was measured by laser scanning confocal fluorescence microscope.RESULTS HepG2 cell viability was significantly reduced following exposure to dictamnine 25-800 μmol·L-1 for 24 h in a concentration-dependent manner(r=0.965,P0.05),and IC50 value was(283±27)μmol·L-1.The LDH release rate of HepG2 cells was significantly increased after exposure to dictamnine 12.5-50 μmol·L-1 for 24 h(P0.01).The morphology of HepG2 cells after 24 h exposure to dictamnine 100 and 200 μmol·L-1 was changed greatly.The cells wrinkled up and dropped.Compared with control group,ALT and AST activities in HepG2 cell culture were significantly increased(P0.05) in a concentration-dependent manner(r=0.995,P0.05 and r=0.996,P0.05) following exposure to dictamnine 100 and 200 μmol·L-1 for 24 h,and the mitochondrial membrane potential markedly declined(r=0.978,P0.05).After exposure to dictamnine 100 and 200 μmol·L-1 for 4 h,GST activity in HepG2 cell culture was significantly increased(P0.05);and for 24 h,GST activity in dictamnine 50,100 and 200 μmol·L-1 groups was also increased in a concentration-dependent manner(r=0.987,P0.05).After exposure to dictamnine 200 μmol·L-1 for 48 h,GGT activity in HepG2 cell culture was increased(P0.05).CONCLUSION Dictamnine at higher concentrations(≥100 μmol·L-1)has potential hepatotoxicity.The cell membrane damage and mitochondrial membrane damage may be involved in the dictamnine-induced hepatotoxity mechanism.
【Fund】: 北京市自然科学基金项目(7122105);; 北京大学985项目(BMU20100119)~~
【CateGory Index】: R285.5
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