体外肝脏3D模型在药物肝毒性评价中的优势

Advantages of in vitro 3D liver model in study of liver toxicity

建立及评价体外肝脏3D模型在药物肝毒性评价中的优势。本研究利用悬滴技术构建Hepa RG3D多细胞聚球体模型,并检测其肝功能水平,最后用该模型评价2个肝毒性阳性药重复给药毒性,并与2D模型下单次给药毒性进行比较。Hepa RG细胞聚集生长形成微组织体,该模型的细胞白蛋白表达水平、尿素分泌水平和CYP3A4活性诱导水平均显著高于2D模型。盐酸胺碘酮在2D和3D培养模型下IC_(50)分别为50和100μmol·L^(-1);异烟肼在2D模型下IC_(50)>1 mmol·L^(-1),3D模型下IC_(50)为700μmol·L^(-1)。两个药物在3D模型下LDH活性均呈现明显的时间/给药次数依赖性和剂量正相关性。结果提示成功建立体外肝脏3D悬滴培养模型,与2D模型比较,该模型可用于准确评价肝毒性阳性药,并为将来体外准确、高通量和多次长期给药评价药物肝脏毒性提供可能。

This study was conducted to establish an in vitro 3D liver model and apply it to the drug liver toxicity evaluation.The 3D multicellular sphere model of Hepa RG cells was established by hanging-drop technique for evaluation of liver function.The 3D liver model was used to test the hepatotoxicity of isoniazid and amiodarone hydrochloride compared to the 2D cell culture model.Our results showed that Hepa RG cells formed a compact spheriod,and the level of cell albumin,urea and the CYP3A4 activity were significantly higher than that of 2D model.With the treatment of amiodarone hydrochloride in 2D and 3D model,the IC（50）were 50 and 100μmol·L^（-1）,respectively.When the dose was less than 1 000μmol·L^（-1）,isoniazid had no hepatocytetoxicity in 2D model,while the IC（50） in 3D model was 700μmol·L^（-1）.The LDH activities of both drugs in 3Dmodel showed time-and dose-dependent correlation.The results suggest that this in vitro 3D hanging-drop liver model is good for testing liver functions with a high hepatic drug-metabolizing enzyme activity.Compared with the 2D model,the 3D liver model can accurately evaluate the liver toxicity of drugs.Our results demonstrated the importance of in vitro cell culture models for detection of in vivo-relevant adverse effects of drugs.