Andersson T, Förlin L, Hansson T
Drug Metab Dispos. 1983 Sep-Oct;11(5):494-8.
A reliable technique for rainbow trout liver perfusion has been developed for studies on xenobiotic biotransformation. Normal function of the perfused liver was indicated throughout the perfusion experiment by 1) a proper oxygen consumption, 2) a low leakage of intracellular enzymes, 3) a stable pH value in the effluent, and 4) a stable release of metabolites into the effluent perfusate and bile for at least 2 hr after maximal rate of metabolism was attained. The main metabolite of 7-ethoxycoumarin in effluent perfusate was identified as 7-hydroxycoumarin glucuronide. Only trace amounts were identified as sulfates. When fish were pretreated with Clophen A50 or beta-naphthoflavone, the amount of metabolites released into the effluent perfusate increased 3.4- and 6.4-fold, respectively, when compared to control livers. Furthermore, in livers from Clophen A50- or beta-naphthoflavone-treated fish, only 80 and 67%, respectively, of excreted products were conjugated. Influence of temperature on 7-ethoxycoumarin metabolism was studied in perfused liver and isolated liver microsomes. Results indicate that the Q10 for the metabolism of 7-ethoxycoumarin in perfused liver deviates from that found in isolated microsomes. The amount of metabolites excreted into the bile consisted of about 25% of the amount found in effluent perfusate. The only metabolite detected in bile from perfused liver from control as well as treated fish was 7-hydroxycoumarin glucuronide.
已开发出一种用于虹鳟鱼肝灌注的可靠技术，用于研究外源生物转化。在整个灌注实验中，灌注肝的正常功能通过以下几点得以体现：1）适当的耗氧量；2）细胞内酶的低泄漏率；3）流出液中稳定的pH值；4）在达到最大代谢率后至少2小时内，代谢产物稳定释放到流出灌注液和胆汁中。流出灌注液中7-乙氧基香豆素的主要代谢产物被鉴定为7-羟基香豆素葡糖醛酸苷。仅检测到痕量的硫酸盐。当用氯芬A50或β-萘黄酮对鱼进行预处理时，与对照肝相比，释放到流出灌注液中的代谢产物量分别增加了3.4倍和6.4倍。此外，在经氯芬A50或β-萘黄酮处理的鱼的肝脏中，分别只有80%和67%的排泄产物被结合。在灌注肝和分离的肝微粒体中研究了温度对7-乙氧基香豆素代谢的影响。结果表明，灌注肝中7-乙氧基香豆素代谢的Q10与分离微粒体中的不同。排泄到胆汁中的代谢产物量约占流出灌注液中发现量的25%。在对照鱼和处理鱼的灌注肝胆汁中检测到的唯一代谢产物是7-羟基香豆素葡糖醛酸苷。
