Di Bello Domenica, Vaccaro Emilia, Longo Vincenzo, Regoli Francesco, Nigro Marco, Benedetti Maura, Gervasi Pier Giovanni, Pretti Carlo
Dipartimento di Patologia Animale Profilassi ed Igiene degli Alimenti, Università di Pisa, Viale delle Piagge 2, 56124 Pisa, Italy.
Aquat Toxicol. 2007 Aug 15;84(1):19-26. doi: 10.1016/j.aquatox.2007.05.010. Epub 2007 May 25.
This study investigated some aspects of xenobiotic metabolism in the Nototheniidae Trematomus bernacchii, a key sentinel species for monitoring Antarctic ecosystems. After laboratory exposure to beta-naphthoflavone (betaNF), basal levels and time-course induction of CYP1A, CYP1B and CYP3A were measured as enzymatic activities, immunoreactive protein content and mRNA expression in liver, gills, intestine and heart. Additional analyses in the liver included enzymatic activities of testosterone hydroxylase, (omega)- and (omega-1)-lauric acid hydroxylase and some phase II enzymes related to the AhR battery genes, DT-diaphorase, glutathione S-transferases and UDP-glucuronyl transferases. Responsiveness of hepatic CYP1A1 after exposure to betaNF demonstrated an higher sensitivity of MEROD than EROD activity and long lasting expression of mRNA still induced after 20 days from the treatment. Testosterone metabolism, oxidation of lauric acid and activities of phase II enzymes were not affected by betaNF indicating that their modulation is not mediated by Ah receptor. Induction of CYP1A was more limited in gills and absent in intestine and heart. The first nucleotide sequence for CYP1B1 in an Antarctic fish has been obtained, revealing a homology of 89% and 72% respectively to CYP1B1 of plaice and CYP1B2 of carp. Constitutive expression of CYP1B1 was restricted to gills where it was also induced by betaNF. Obtained results represent an additional contribution to the ecotoxicological characterization of T. bernacchii and further support the use of biomarkers for early detection of chemical pollution in Antarctica.
本研究调查了南极生态系统监测关键指示物种——南极鱼科的伯氏南极鱼(Trematomus bernacchii)对外源物质代谢的某些方面。在实验室中将其暴露于β-萘黄酮(βNF)后,测定了肝脏、鳃、肠道和心脏中CYP1A、CYP1B和CYP3A的基础水平以及随时间变化的诱导情况,包括酶活性、免疫反应性蛋白含量和mRNA表达。肝脏中的其他分析包括睾酮羟化酶、ω-和ω-1-月桂酸羟化酶以及一些与芳烃受体(AhR)相关基因、DT-黄递酶、谷胱甘肽S-转移酶和UDP-葡萄糖醛酸基转移酶相关的II相酶的酶活性。暴露于βNF后肝脏CYP1A1的反应性表明,甲氧基异吩恶唑脱甲基酶(MEROD)活性比乙氧基异吩恶唑脱甲基酶(EROD)活性具有更高的敏感性,并且在处理后20天仍诱导mRNA的持久表达。睾酮代谢、月桂酸氧化和II相酶活性不受βNF影响,表明它们的调节不是由Ah受体介导的。CYP1A在鳃中的诱导作用更有限,在肠道和心脏中未出现诱导。获得了南极鱼类中CYP1B1的首个核苷酸序列,分别显示与鲽鱼的CYP1B1和鲤鱼的CYP1B2具有89%和72%的同源性。CYP1B1的组成型表达仅限于鳃,在鳃中它也被βNF诱导。所得结果为伯氏南极鱼的生态毒理学特征提供了额外贡献,并进一步支持使用生物标志物早期检测南极的化学污染。