摘要
目的观察酮康唑(ketoconazole,KTZ)对人L02肝细胞的毒性效应,鉴定细胞毒性差异蛋白。方法KTZ浓度为30、50、80和100mg/L,与人L02肝细胞共培养2、4、6和8h,采用CCK-8法和流式细胞术检测KTZ对细胞增殖和胞内活性氧的影响;提取产生明显毒性的实验组细胞和对照组细胞的胞内蛋白质,进行双向电泳,并用质谱鉴定差异性蛋白。结果细胞增殖抑制率呈剂量依赖性显著性升高,胞内活性氧呈剂量依赖性显著性升高。共鉴定出7种有功能的细胞内差异性蛋白。结论鉴定出的差异性蛋白为KTZ毒性机制研究提供新的参考。
Objective To observe the toxicity effects of Ketoconazole on human L02 hepatocytes,and identify the intracellular toxicity differential proteins.Methods Human L02 hepatocytes were co-cultured with 30,50,80 and 100mg/L of ketoconazole for 2,4,6 and 8h,CCK-8 assay and flow cytometry were used to determine the cell viability and the intracellular reactive oxygen species(ROS)after Ketoconazole treatment;Intracellular proteins of test group when toxicity occurred and control group were extracted,and the proteins were identified by two-dimensional electrophoresis(2D-PAGE)and liquid chromatography-tandem mass spectrometry(LC-MS/MS).Results The cell proliferation inhibition rate and the intracellular ROS increased significantly in a dose-dependent manner by Ketoconazole treatment.Seven intracellular differential proteins in test group were identified.Conclusion The differential proteins identified can provide a new reference for the study of the toxicity mechanism of ketoconazole.
作者
苗玉发
康慧君
王晓姝
李路路
张河战
Miao Yufa;Kang Huijun;Wang Xiaoshu(National Institutes for Food and Drug Control,Beijing 100176,China)
出处
《医学研究杂志》
2019年第9期37-40,44,共5页
Journal of Medical Research
基金
国家“重大新药创制”科技重大专项基金资助项目(2018ZX09201017)
