应用高内涵分析技术进行药物肝毒性风险评估的研究  

Study on the risk assessment of drug hepatoxicty by high content analysis

作　　者：高阳[1] 胡宇驰[1] 左泽平[1] 王志斌[1] 金家金 王立娜[2] 程源[2] 

机构地区：[1]中药成分分析与生物评价北京市重点实验室北京市保健食品化妆品检验中心北京市药品检验所,北京100035 [2]北京中医药大学,北京100102

出　　处：《药学研究》2015年第12期688-691,702,共5页Journal of Pharmaceutical Research

基　　金：2013年北京市科技计划项目(No.Z131100005613032)

摘　　要：目的研究尝试应用高内涵分析技术建立药物肝毒性高内涵毒性评价指数测定的标准方法。方法采用Tox Insight药物肝毒性检测试剂盒,利用对多靶点的特异性染色,结合高内涵活细胞成像分析平台考察4种药物对人源性肝癌细胞Hep G2的细胞毒性作用,包括对该细胞的细胞数、核DNA含量、细胞质内谷胱甘肽降低水平、活性氧水平、线粒体膜电位的改变等5个指标进行综合分析,拟对受试样品的肝毒性进行评估。结果氟西汀和罗格列酮的肝毒性评估结果为阴性;甲芬那酸和噻氯匹定的肝毒性评估结果为阳性,提示具有肝毒性风险,且甲芬那酸的肝毒性风险比噻氯匹定的高。结论应用高内涵分析技术评估药物肝毒性的方法基于活细胞、多参数、实时、高通量,能够实现药物多种生物活性、毒性的早期、快速检测,并且灵敏稳定,简便可行,可尝试用于对药物进行体外肝毒性的安全风险评估。Objective To study a standard method for the determination of drug hepatoxicty evaluation index using high content analysis. Methods To investigate the cytotoxic effect of four drugs on HepG2 cells in human hepatocellular carcinoma cells by using ToxInsight Drug Induced Liver Injury Assay Cartridge and combining high content live cell imaging analysis platform. Through the comprehensive analysis of 5 indexes,the number of cells,DNA content,GSH level,ROS level and mitochondrial membrane potential,the liver toxicity of the tested samples were evaluated. Results The results of fluox-etine and rosiglitazone were negative. The results of mefenamic acid and ticlopidine were positive,which indicated that the risk of liver toxicity was high,and the risk of liver toxicity of mefenamic acid was higher than that of ticlopidine. Conclusion HCA was based on living cells,multi parameters,real - time and high content,can achieve a variety of biological activity, drug toxicity of the early and rapid detection. It was sensitive,stable,simple and feasible. HCA can be used to evaluate the risk of drug hepatoxicty in vitro.

关 键 词：高内涵分析 药物肝毒性 毒性评估 

分 类 号：R965[医药卫生—药理学]