Stegeman J J, Woodin B R, Singh H, Oleksiak M F, Celander M
Biology Department, Woods Hole Oceanographic Institution, MA 02543, USA.
Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1997 Jan;116(1):61-75. doi: 10.1016/s0742-8413(96)00128-4.
Hepatic microsomes prepared from 10 fish species from Bermuda were studied to establish features of cytochrome P450 (CYP) systems in tropical marine fish. The majority (7/10) of the species had total P450 content between 0.1 and 0.5 nmol/mg, and cytochrome b5 content between 0.025 and 0.25 nmol/mg. Ethoxycoumarin O-deethylase (ECOD) and aminopyrine N-demethylase (APND) rates in these 7 species were 0.23-2.1 nmol/min/mg and 0.5-11 nmol/min/mg, respectively, similar to rates in many temperate fish species. In contrast to those 7 species, sergeant major (Abudefduf saxatilis) and Bermuda chub (Kyphosus sectatrix) had microsomal P450 contents near 1.7 nmol/mg, among the highest values reported in untreated fish, and had greater rates of ECOD, APND, ethoxyresorufin O-deethylase (EROD) and pentoxyresorufin O-depentylase than did most of the other species. Freshly caught individuals of all species had detectable levels of EROD and aryl hydrocarbon hydroxylase (AHH) activities. Those individuals with higher rates of EROD activity had greater content of immunodetected CYP1A protein, consistent with Ah-receptor agonists acting to induce CYP1A in many fish in Bermuda waters. Injection of tomtate and blue-striped grunt with beta-naphthoflavone (BNF; 50 or 100 mg/kg) induced EROD rates by 25 to 55-fold, suggesting that environmental induction in some fish was slight compared with the capacity to respond. AHH rates were induced only 3-fold in these same fish. The basis for disparity in the degree of EROD and AHH induction is not known. Rates of APND and testosterone 6 beta- and 16 beta-hydroxylase were little changed by BNF, indicating that these are not CYP1A activities in these fish. Antibodies to phenobarbital-inducible rat CYP2B1 or to scup P450B, a putative CYP2B, detected one or more proteins in several species, suggesting that CYP2B-like proteins are highly expressed in some tropical fishes. Generally, species with greater amounts of total P450 had greater amounts of proteins related to CYP2B. These species also had appreciable amounts of CYP3A-like proteins. Thus, many fishes in Bermuda appear to have induced levels of CYP1A; some also have unusually high levels of total P450 and of CYP2B-like and CYP3A-like proteins. These species may be good models for examining the structural, functional and regulatory properties of teleost CYP and the environmental or ecological factors contributing to high levels of expression of CYP in some fishes.
对从百慕大群岛的10种鱼类制备的肝微粒体进行了研究,以确定热带海洋鱼类细胞色素P450(CYP)系统的特征。大多数(7/10)物种的总P450含量在0.1至0.5 nmol/mg之间,细胞色素b5含量在0.025至0.25 nmol/mg之间。这7个物种中乙氧香豆素O - 脱乙基酶(ECOD)和氨基比林N - 脱甲基酶(APND)的活性分别为0.23 - 2.1 nmol/min/mg和0.5 - 11 nmol/min/mg,与许多温带鱼类的活性相似。与这7个物种不同,军士长鱼(Abudefduf saxatilis)和百慕大鲹(Kyphosus sectatrix)的微粒体P450含量接近1.7 nmol/mg,是未处理鱼类中报道的最高值之一,并且其ECOD、APND、乙氧试卤灵O - 脱乙基酶(EROD)和戊氧试卤灵O - 脱戊基酶的活性比大多数其他物种更高。所有物种的新鲜捕获个体均具有可检测水平的EROD和芳烃羟化酶(AHH)活性。那些具有较高EROD活性的个体,其免疫检测到的CYP1A蛋白含量更高,这与在百慕大水域许多鱼类中Ah受体激动剂诱导CYP1A的作用一致。给汤氏魣和蓝条魮注射β - 萘黄酮(BNF;50或100 mg/kg)可使EROD活性提高25至55倍,这表明与某些鱼类的反应能力相比,环境诱导作用在某些鱼类中较小。在这些相同的鱼类中,AHH活性仅诱导了3倍。EROD和AHH诱导程度差异的原因尚不清楚。BNF对APND以及睾酮6β - 和16β - 羟化酶的活性影响很小,表明这些不是这些鱼类中的CYP1A活性。针对苯巴比妥诱导的大鼠CYP2B1或鲷鱼P450B(一种假定的CYP2B)的抗体在几个物种中检测到一种或多种蛋白质,这表明CYP2B样蛋白在一些热带鱼类中高度表达。一般来说,总P450含量较高的物种,与CYP2B相关的蛋白质含量也较高。这些物种还具有相当数量的CYP3A样蛋白。因此,百慕大的许多鱼类似乎具有诱导水平的CYP1A;一些鱼类还具有异常高的总P450以及CYP2B样和CYP3A样蛋白水平。这些物种可能是研究硬骨鱼CYP的结构、功能和调节特性以及导致某些鱼类中CYP高表达的环境或生态因素的良好模型。