Uehara Shotaro, Higuchi Yuichiro, Yoneda Nao, Yamazaki Hiroshi, Suemizu Hiroshi
a Central Institute for Experimental Animals , Kawasaki , Japan.
b Laboratory of Drug Metabolism and Pharmacokinetics , Showa Pharmaceutical University , Machida , Tokyo , Japan.
Xenobiotica. 2019 Jun;49(6):678-687. doi: 10.1080/00498254.2018.1495346. Epub 2018 Oct 4.
The evaluation of drug-mediated cytochrome P450 (P450) induction using human hepatocytes is important for predicting drug interactions. In this study, we prepared hepatocytes from chimeric mice with humanised livers (Hu-Liver mice) and evaluated the expression and inducibility of P450s in these hepatocytes. Up to 95% of the Hu-Liver cells stained positive for human leukocyte antigen and the mean viability exceeded 85% (n = 10). Monolayer-cultured Hu-Liver cells displayed a similar morphology to cultures of the corresponding human hepatocytes used as transplantation donors. The mRNA expression levels in Hu-Liver cells of 16 P450 forms belonging to P450 subfamilies 1-4 correlated well with the expression levels of the same enzymes in human hepatocytes. The variations in individual P450 mRNA levels between Hu-Liver cells and the corresponding human hepatocytes were within five-fold for 13 P450 forms. The production of 6β-hydroxytestosterone in Hu-Liver cells was significantly increased (p < .05) following treatment with the CYP3A inducer, rifampicin. Hu-Liver cells have characteristics similar to those of human hepatocytes in terms of mRNA expression levels and the inducibility of the various P450 forms. Thus, Hu-Liver cells can potentially be used for in vitro drug-mediated induction assays of human hepatic P450s.
利用人肝细胞评估药物介导的细胞色素P450（P450）诱导作用对于预测药物相互作用很重要。在本研究中，我们从具有人源化肝脏的嵌合小鼠（人源化肝脏小鼠）制备了肝细胞，并评估了这些肝细胞中P450的表达和诱导性。高达95%的人源化肝脏细胞人白细胞抗原染色呈阳性，平均活力超过85%（n = 10）。单层培养的人源化肝脏细胞呈现出与用作移植供体的相应人肝细胞培养物相似的形态。属于P450亚家族1 - 4的16种P450形式在人源化肝脏细胞中的mRNA表达水平与在人肝细胞中相同酶的表达水平高度相关。对于13种P450形式，人源化肝脏细胞与相应人肝细胞之间的个体P450 mRNA水平差异在五倍以内。用CYP3A诱导剂利福平处理后人源化肝脏细胞中6β - 羟基睾酮的产生显著增加（p < 0.05）。就mRNA表达水平和各种P450形式的诱导性而言，人源化肝脏细胞具有与人肝细胞相似的特征。因此，人源化肝脏细胞有可能用于人肝P450的体外药物介导诱导试验。
