University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Zatisi 728/II, 389 25 Vodnany, Czech Republic.
Swedish University of Agricultural Sciences, Department of Molecular Science, Uppsala BioCenter, P.O. Box 7015, SE-750 07 Uppsala, Sweden.
Chemosphere. 2018 Aug;205:380-386. doi: 10.1016/j.chemosphere.2018.04.080. Epub 2018 Apr 14.
This study examined the ability of several human pharmaceuticals to modulate hepatic piscine CYP-mediated monooxygenase activities. Effects of six pharmaceuticals: diclofenac, sulfamethoxazole, tramadol, carbamazepine, venlafaxine and nefazodone, were investigated in vitro in rainbow trout hepatic microsomes. The reactions of 7-ethoxyresorufin-O-deethylase (EROD) and benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylase (BFCOD), were used as markers for hepatic CYP1A and CYP3A-like activities, respectively. Our results showed that EROD and BFCOD activities were both affected by nefazodone. Nefazodone inhibited EROD in a dose dependent manner and was found to be a potent non-competitive inhibitor of EROD with a K value of 6.6 μM. BFCOD activity was inhibited non-competitively in the presence of nefazadone with K value of 30.7 μM. BFCOD activity was slightly reduced only by the highest concentration of carbamazepine. Diclofenac, sulfamethoxazole, tramadol, and venlafaxine did not affect the activity of either EROD or BFCOD. We further exposed microsomal fraction to mixtures of six pharmaceuticals to investigate potential inhibition. The results showed that EROD and BFCOD activity was inhibited on 94% and 80%, respectively at higher tested concentration. To our knowledge, this is the first report to demonstrate an inhibitory effect of nefazodone on hepatic CYP1A and CYP3A-like proteins in rainbow trout.
本研究考察了几种人用药物对鱼类 CYP 介导的单加氧酶活性的调节能力。在虹鳟鱼肝微粒体中，研究了六种药物（双氯芬酸、磺胺甲恶唑、曲马多、卡马西平、文拉法辛和奈法唑酮）的体外作用。7-乙氧基resorufin-O-脱乙基酶（EROD）和苄氧基-4-三氟甲基香豆素-O-去苄氧基酶（BFCOD）的反应分别作为肝 CYP1A 和 CYP3A 样活性的标志物。我们的结果表明，EROD 和 BFCOD 活性均受奈法唑酮的影响。奈法唑酮呈剂量依赖性抑制 EROD，是 EROD 的强非竞争性抑制剂，K 值为 6.6 μM。在奈法唑酮存在下，BFCOD 活性呈非竞争性抑制，K 值为 30.7 μM。只有最高浓度的卡马西平才会轻微降低 BFCOD 活性。双氯芬酸、磺胺甲恶唑、曲马多和文拉法辛均不影响 EROD 或 BFCOD 的活性。我们进一步将微粒体部分暴露于六种药物混合物中，以研究潜在的抑制作用。结果表明，在较高的测试浓度下，EROD 和 BFCOD 活性分别抑制了 94%和 80%。据我们所知，这是首次报道奈法唑酮对虹鳟鱼肝 CYP1A 和 CYP3A 样蛋白的抑制作用。
