Baron Matthew G, Mintram Kate S, Owen Stewart F, Hetheridge Malcolm J, Moody A John, Purcell Wendy M, Jackson Simon K, Jha Awadhesh N
School of Biological Science, Plymouth University, Devon, United Kingdom.
AstraZeneca, Alderley Park, Macclesfield, Cheshire, United Kingdom.
PLoS One. 2017 Jan 3;12(1):e0168837. doi: 10.1371/journal.pone.0168837. eCollection 2017.
At high internal doses, pharmaceuticals have the potential for inducing biological/pharmacological effects in fish. One particular concern for the environment is their potential to bioaccumulate and reach pharmacological levels; the study of these implications for environmental risk assessment has therefore gained increasing attention. To avoid unnecessary testing on animals, in vitro methods for assessment of xenobiotic metabolism could aid in the ecotoxicological evaluation. Here we report the use of a 3-D in vitro liver organoid culture system (spheroids) derived from rainbow trout to measure the metabolism of seven pharmaceuticals using a substrate depletion assay. Of the pharmaceuticals tested, propranolol, diclofenac and phenylbutazone were metabolised by trout liver spheroids; atenolol, metoprolol, diazepam and carbamazepine were not. Substrate depletion kinetics data was used to estimate intrinsic hepatic clearance by this spheroid model, which was similar for diclofenac and approximately 5 fold higher for propranolol when compared to trout liver microsomal fraction (S9) data. These results suggest that liver spheroids could be used as a relevant and metabolically competent in vitro model with which to measure the biotransformation of pharmaceuticals in fish; and propranolol acts as a reproducible positive control.
在高剂量内服时，药物有可能在鱼类中诱导产生生物学/药理学效应。对环境而言，一个特别令人担忧的问题是它们具有生物累积并达到药理水平的可能性；因此，对这些影响进行环境风险评估的研究受到了越来越多的关注。为避免对动物进行不必要的试验，用于评估外源化合物代谢的体外方法有助于生态毒理学评价。在此，我们报告了使用源自虹鳟鱼的三维体外肝脏类器官培养系统（球体），通过底物消耗试验来测量七种药物的代谢情况。在所测试的药物中，普萘洛尔、双氯芬酸和苯基布他松被虹鳟鱼肝脏球体代谢；阿替洛尔、美托洛尔、地西泮和卡马西平则未被代谢。利用底物消耗动力学数据通过该球体模型估算肝脏固有清除率，与虹鳟鱼肝脏微粒体组分（S9）数据相比，双氯芬酸的结果相似，而普萘洛尔的结果约高5倍。这些结果表明，肝脏球体可作为一种相关且具有代谢活性的体外模型，用于测量鱼类中药物的生物转化；并且普萘洛尔可作为可重复的阳性对照。
