In vitro modulation of 17-beta-estradiol-induced vitellogenin synthesis: Effects of cytochrome P4501A1 inducing compounds on rainbow trout (Oncorhynchus mykiss) liver cells
In vitro modulation of 17-beta-estradiol-induced vitellogenin synthesis: Effects of cytochrome P4501A1 inducing compounds on rainbow trout (Oncorhynchus mykiss) liver cells
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DOI：
10.1016/0166-445x(95)00047-8
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发表时间：
1996-04-01
影响因子：
4.5
通讯作者：
Hinton, DE
中科院分区：
文献类型：
--
作者：
Anderson, MJ;Miller, MR;Hinton, DE
细胞色素P4501A1(CYP1A1)的诱导，特别是在肝脏中，是鱼类暴露于多环和卤代芳香烃的一个有用的标志。然而，鱼类细胞色素P1A1的诱导与毒性之间的关系尚不确定。在哺乳动物生物实验中，一些诱导CYP1A1的化合物是抗雌激素的，这种作用与芳香烃(Ah)受体和/或增加17-β-雌二醇的分解代谢有关。鱼类肝脏合成雌激素诱导的卵黄前体蛋白卵黄蛋白原(VG)，对卵母细胞成熟和卵巢发育至关重要。为了确定在鱼类肝脏中是否存在与细胞色素P1A1相关的内分泌调节，将17-β-雌二醇和细胞色素P1A1诱导的化合物或混合物共同作用于原代培养的虹鳟鱼肝细胞。在15℃的改良HEPES缓冲培养液199中培养细胞的蛋白质合成和酶活性最好，Vg和白蛋白(Alb)是试验的基础，通过EL ISA测量处理后48h的浓度估计白蛋白(Alb)。评估相当的存活率(线粒体脱氢酶活性)和分泌功能(Alb合成)，并与其他结果相关联。2，3，4，7，8-五氯二苯并呋喃(10(-12)~10(-8)M)和GT；与2，3，7，8-四氯二苯并对二恶英(TCDD)相同的2，3，7，8-四氯二苯并呋喃(10(-12)~10(-8)M)和GT；β-萘黄酮(10(-7)~10(-6)M)抑制17-β-雌二醇处理的肝细胞VG合成。抑制作用的强弱与其作为CYP1A1蛋白诱导剂的强度直接相关。在10(-8)M时，多氯联苯同系物77、126、156不抑制VG的合成，并诱导NO至中等水平的CYP1A1蛋白或EROD活性。在10(-8)M时，同系物114，一种弱的EROD诱导剂，相对于单独用17-β-雌二醇处理的细胞，增强了VG的合成。这项研究的结果增加了我们对肝脏细胞色素P1A1诱导的后果的理解，为性成熟鱼类暴露于细胞色素P1A1诱导化合物的生殖损害提供了预警，并为进一步的更详细的体内研究提供了依据。
Cytochrome P4501A1 (CYP1A1) induction, particularly in liver, is a useful marker of exposure of fish to polycyclic- and halogenated-aromatic hydrocarbons. However, the relationship between toxicity and CYP1A1 induction in fish is uncertain. Some compounds that induce CYP1A1 are antiestrogenic in mammalian bioassay, and this effect is linked to aryl hydrocarbon (Ah) receptor and/or increased catabolism of 17-beta-estradiol. Liver of fish synthesizes estrogen-inducible egg yolk precursor protein vitellogenin (Vg) which is critical for oocyte maturation and ovarian development. To determine if CYP1A1-associated endocrine modulation could occur in fish liver, primary cultures of rainbow trout liver cells were co-administered 17-beta-estradiol and CYP1A1 inducing compounds or mixtures. Protein synthesis and enzyme activity of cells were optimal when cultured in a modified HEPES buffered Medium 199 at 15 degrees C. Vg and albumin (Alb), estimated by ELISA measurement of concentration in the media 48 h after treatment, formed the basis for the test. Equivalent viability (mitochondrial dehydrogenase activity) and secretory functional capacity (Alb synthesis) were estimated and correlated with other results. In descending order, 2,3,4,7,8-pentachlorodibenzofuran (10(-12) to 10(-8) M) > 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) congruent to 2,3,7,8-tetrachlorodibenzofuran (10(-12) to 10(-8) M) > beta-naphthoflavone (10(-7) to 10(-6) M) inhibited Vg synthesis in 17-beta-estradiol treated liver cells. Potency of inhibition directly related to strength as an inducer of CYP1A1 protein. At 10(-8) M, PCB congeners 77, 126, 156 did not inhibit Vg synthesis and induced no to moderate levels of CYP1A1 protein or EROD activity. At 10(-8) M, congener 114, a weak EROD inducer, potentiated Vg synthesis relative to cells treated with 17-beta-estradiol alone. The results of this study increase our understanding of the consequences of hepatic CYP1A1 induction, forewarn of reproductive impairment of sexually maturing fishes exposed to CYP1A1 inducing compounds and argue for further, more detailed in vivo investigation.