Synergistic induction of drug-metabolizing enzymes in co-cultures of bovine hepatocytic and sinusoidal cell lines
Synergistic induction of drug-metabolizing enzymes in co-cultures of bovine hepatocytic and sinusoidal cell lines
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牛肝细胞和正弦细胞系共培养物中药物代谢酶的协同诱导
DOI：
10.1007/s11626-019-00408-6
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Yamanaka N
中科院分区：
文献类型：
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作者：
Yoshioka M;Takenouchi T;Kitani H;Guruge KS;Yamanaka N
肝细胞来源的细胞系为研究肝脏代谢提供了有用的实验系统。与人类和啮齿类动物不同，很少有肝细胞来源的细胞系是从牛身上产生的。在这里，我们通过转染SV40大t抗原构建体建立了两种永活牛肝细胞来源的细胞系（BH4和BH5）。形态学和免疫细胞化学分析显示BH4和BH5分别来源于肝细胞和胆道上皮细胞。一种强致癌物，3-甲基胆蒽（3-MC），在BH4和BH5中上调细胞色素P450 (CYP)1A1、CYP1A2和CYP1B1的基因表达，但其水平不到原代培养牛肝细胞的十五分之一。苯巴比妥（PB）也增加了BH4和BH中CYP2B6、CYP2C18和CYP3A4的表达水平，但低于3-MC。相比之下，当BH4或BH5与先前建立的牛肝血管细胞系共培养并经3-MC处理时，与单独培养的BH4或BH5细胞相比，CYP1A1、CYP1A2和CYP1B1的基因表达量增加了38~290%。铅处理的BH4或BH5细胞与肝窦细胞系共培养也显示出CYP2B6和CYP2C18表达的协同增加。综上所述，这些共培养物可为研究药物在牛肝脏中的药理和毒理学特性提供动物模型。
Hepatocyte-derived cell lines provide useful experimental systems for studying liver metabolism. Unlike human and rodents, few hepatocyte-derived cell lines have been generated from cattle. Here, we established two immortalized bovine hepatocyte-derived cell lines (BH4 and BH5) via transfection with a SV40 large T-antigen construct. Morphological and immunocytochemical analyses revealed that BH4 and BH5 originated from hepatocytes and biliary-epithelial cells, respectively. A potent carcinogen, 3-methylcholanthrene (3-MC), upregulated gene expression of cytochrome P450 (CYP)1A1, CYP1A2, and CYP1B1 in BH4 and BH5, but only to levels less than one-fifteenth of those in primary cultured bovine hepatocytes. Phenobarbital (PB) also increased expression levels of CYP2B6, CYP2C18, and CYP3A4 in BH4 and BH, but at a lower level than 3-MC. By contrast, when BH4 or BH5 was co-cultured with previously established bovine liver sinusoidal cell lines and treated with 3-MC, the gene expression levels of CYP1A1, CYP1A2, and CYP1B1 increased by 38~290%, compared with those in BH4 or BH5 cells cultured alone. PB-treated co-cultures of BH4 or BH5 cells and liver sinusoidal cell lines also showed synergistic increases in CYP2B6 and CYP2C18 expression. Together, the results suggest that these co-cultures could provide anin vitromodel for investigations into pharmacological and toxicological properties of drugs in cattle liver.