Effects of steatosis on drug-metabolizing capability of primary human hepatocytes
Effects of steatosis on drug-metabolizing capability of primary human hepatocytes
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DOI：
10.1016/j.tiv.2006.07.008
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发表时间：
2007-03-01
影响因子：
3.2
通讯作者：
Gomez-Lechon, M. J.
中科院分区：
文献类型：
--
作者：
Donato, M. T.;Jimenez, N.;Gomez-Lechon, M. J.
由于大脂肪变性而放弃移植的肝移植物是否适合用于制备体外药物代谢研究的人肝细胞培养物。与正常组织相比，脂肪肝（bbb40 %脂肪变性）的细胞活力和分离率较低。在脂肪肝制备的肝细胞中发现7-乙氧基香豆素o -去乙基化（ECOD）和睾酮氧化显著减少。通过长链游离脂肪酸（FFA）体外培养肝细胞，研究脂质积累对P450酶的潜在影响。0.25-3 mM FFA处理细胞可诱导细胞质中脂质的剂量依赖性积累。暴露于1 mM或2 mM FFA（分别约为对照组的60-70%和30-60%）14小时后，发现ECOD和睾酮氧化降低。从活性和mRNA水平分析脂肪超载对个体p450的影响。肝细胞与I mM（对照组的45-65%）或2 mM（对照组的20-50%）FFA孵育14小时后，CYP1A2、CYP2C9、CYP2E1和CYP3A4活性降低。在I mM FFA处理的肝细胞中也发现P450转录物的减少。我们的研究结果显示，在脂肪超载的肝细胞中，参与药物代谢的p450普遍下调。结果表明，尽管P450功能降低，但从脂肪变性供体获得的人肝细胞具有代谢能力，可用于药物代谢研究。(c) 2006 Elsevier Ltd.版权所有。
The suitability of liver grafts discarded for transplantation because of macrosteatosis for preparing human hepatocyte cultures for in vitro drug metabolism studies has been examined. Lower cell viability and yield of isolation procedure were obtained from fatty livers (> 40% steatosis) with respect to normal tissue. Significant reductions in 7-ethoxycoumarin O-deethylation (ECOD) and testosterone oxidations were found in hepatocytes prepared from steatotic livers. The potential impact of lipid accumulation on P450 enzymes was studied in vitro by incubation of cultured hepatocytes with long chain free fatty acids (FFA). Treatment of cells with 0.25-3 mM FFA induced dose-dependent accumulation of lipids in the cytosol. Decreased ECOD and testosterone oxidation were found after 14 h of exposure to I mM or 2 mM FFA (about 60-70% and 30-60% of control, respectively). The effects of fat-overloading on individual P450s were analyzed both at activity and mRNA level. CYP1A2, CYP2C9, CYP2E1 and CYP3A4 activities were reduced after hepatocyte incubation with I mM (to 45-65% of control) or 2 mM (to 20-50%) FFA for 14 h. Reductions in P450 transcripts were also found in hepatocytes treated with I mM FFA. Our findings showed a general down-regulation of P450s involved in drug metabolism in fat-overloaded hepatocytes. The results suggest that, despite their reduced P450 function, human hepatocytes obtained from donors with steatosis are metabolically competent and could be used for drug metabolism studies. (c) 2006 Elsevier Ltd. All rights reserved.