Cytochrome P450 isozyme protein verified in the skin of southern hemisphere humpback whales (Megaptera novaeangliae): implications for biochemical biomarker assessment

Large mysticete whales represent a unique challenge for chemical risk assessment. Few epidemiological investigations are possible due to the low incidence of adult stranding events. Similarly their often extreme life-history adaptations of prolonged migration and fasting challenge exposure assumptions. Molecular biomarkers offer the potential to complement information yielded through tissue chemical analysis, as well as providing evidence of a molecular response to chemical exposure. In this study we confirm the presence of cytochrome P450 reductase (CPR) and cytochrome P450 isoenzyme 1A1 (CYP1A1) in epidermal tissue of southern hemisphere humpback whales (Megaptera novaeangliae). The detection of CYP1A1 in the integument of the humpback whale affords the opportunity for further quantitative non-destructive investigations of enzyme activity as a function of chemical stress.     

黄芩苷在中国对虾体内对诺氟沙星消除及细胞色素P450 酶的影响

对照组中国对虾(Fenneropenaeus chinensis)连续投喂不含药物饵料,诺氟沙星组和联合用药组以50 mg/kg体质量剂量连续投喂含诺氟沙星药饵5 d后,诺氟沙星组投喂不含药物饵料10 d,联合用药组以100 mg/kg体质量剂量投喂含黄芩苷饵料10 d。从投喂诺氟沙星药饵时开始,在不同时间点采集中国对虾血液、肝胰腺、鳃和肌肉组织进行药物残留和肝药酶活性测定。结果表明:与诺氟沙星组相比,联合用药组各组织中诺氟沙星消除较快,血淋巴、肝胰腺、鳃和肌肉的理论休药期比诺氟沙星组分别缩短了23.92%、22.73%、25.92%和21.92%;与对照组相比,诺氟沙星对中国对虾CYP1A(ECOD)和CYP2(APND)酶活性有一定的抑制作用,但随着诺氟沙星的消除抑制作用逐渐减弱,最后恢复至对照水平;黄芩苷在加速诺氟沙星在中国对虾体内消除的同时,对中国对虾CYP450酶也有较强的诱导作用。因此,药物使用应注意药物之间的相互作用,以免造成药物中毒或疗效下降现象。     

PCB exposure in sea otters and harlequin ducks in relation to history of contamination by the Exxon Valdez oil spill

Exposure to contaminants other than petroleum hydrocarbons could confound interpretation of Exxon Valdez oil spill effects on biota at Prince William Sound, Alaska. Hence, we investigated polychlorinated biphenyls (PCBs) in blood of sea otters and harlequin ducks sampled during 1998. PCB concentrations characterized by lower chlorinated congeners were highest in sea otters from the unoiled area, whereas concentrations were similar among harlequin ducks from the oiled and unoiled area. Blood enzymes often elevated by xenobiotics were not related to PCB concentrations in sea otters. Only sea otters from the unoiled area had estimated risk from PCBs, and PCB composition or concentrations did not correspond to reported lower measures of population performance in sea otters or harlequin ducks from the oiled area. PCBs probably did not influence limited sea otter or harlequin duck recovery in the oiled area a decade after the spill.     

大黄鱼(Pseudosciaena crocea)细胞色素 P450 CYP4F7基因的克隆及表达分析

CYP4F7是细胞色素P450超基因家族中CYP4F亚家族中的同工酶之一,而细胞色素P450在通过电子传递参与生物体代谢和转化内源和外源化合物方面起着重要的作用。以本实验室构建的大黄鱼消减杂交cDNA文库中623bp的CYP4F7片段设计引物,以大黄鱼的总RNA为模板,利用RACE-PCR的方法,克隆鉴定出了大黄鱼细胞色素P450 CYP4F7基因。结果表明:基因全长1934bp,5'端非编码区59bp,3'端非编码区264bp,开放阅读框1611bp,编码536个氨基酸。序列分析表明大黄鱼的CYP4F7氨基酸序列与舌齿鲈的相似性为91%。利用RT-PCR方法研究CYP4F7在大黄鱼各组织中的表达特征,结果表明CYP4F7在肝脏、脾脏中表达量最高,在心脏、肾脏、头肾、鳃丝中表达量次之。另外,CYP4F7在注射了细菌脂多糖的大黄鱼各组织中的表达量均低于正常的大黄鱼,认为细菌脂多糖可能是CYP4F7表达的抑制剂,说明CYP4F7可能与鱼类免疫反应有一定相关性。     

Cloning of cytochrome P450 1A (CYP1A) genes from the hermaphrodite fish Rivulus marmoratus and the Japanese medaka Oryzias latipes

To use two small fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) and the Japanese medaka Oryzias latipes (Belloniformes) as testing models in molecular ecotoxicology, we have cloned the cytochrome P450 1A (CYP1A) gene after screening of both genomic DNA libraries, and sequenced 11,863 and 7,243 bp including all the exons and introns with promoter regions, respectively. The Rivulus and the medaka CYP1A gene consisted of seven exons (including non-coding exons) with high homology to mammals. In the promoter region, Rivulus CYP1A gene has seven xenobiotic response elements (XREs) and two metal response elements (MREs), while the Japanese medaka CYP1A gene has six XREs and four MREs. Interestingly, medaka CYP1A gene has a number of MREs at the promoter, which may affect its response on metal exposure. We describe here the gene structure of both fish CYP1A genes.     

Inhibitory effects of divalent metal ions on liver microsomal 7-ethoxyresorufin O-deethylase (EROD) activity of leaping mullet

The purpose of the present study was to elucidate in vitro effects of Hg²⁺, Zn²⁺, Ni²⁺ and Cd²⁺ on cytochrome P4501A1 (CYP1A1) dependent EROD activities in leaping mullet liver microsomes. Fish captured from the most polluted part of Izmir Bay, had highly elevated EROD activities, and induced CYP1A1 protein levels as determined by Western blotting. Although all of the metal ions caused inhibition of the initial velocity of the reaction, Hg²⁺ and Cd²⁺ exhibited much higher inhibitory effect at lower concentrations and they were evidently more potent inhibitors than others. The inhibitor concentration giving 50% inhibition (IC₅₀ values) of Zn²⁺, Ni²⁺, Cd²⁺ and Hg²⁺ of initial EROD activity were 107, 16, 1.3 and 0.15 micromolar, respectively. Glutathione (GSH) at 0.5 mM final concentration, completely reversed Ni²⁺ and Cd²⁺ inhibition of EROD activity indicating the protective action of GSH.     

Does CYP1A1polymophism increase the risk for cancer in pollution exposure? A study by gene ontology

Many genetic polymorphisms in metabolism enzymes are important for the risk of cancer as shown in a large number of case-control studies. Cytochrome P-450 (CYP) 1A1 polymorphism is a widely mentioned for importance of its genetic polymorphisms in risk assessment. However, the reports on the correlation between polymorphism and risk are inconsistent. To study the functional aberration in the human wild and mutate types of CYP1A1 is hard. Here, the author used a new gene ontology technology to predict the molecular function of human CYP1A1. Here, it can be seen that there is no functional difference between wild and mutate types of CYP1A1. Therefore, there should be no difference in effect of wild and mutate types of CYP1A1. This can support the null effect of this polymorphism in clinical findings of the population. This polymorphism might not be an important tool for risk assessment.     

黄芩苷对中国对虾细胞色素P450酶及谷光甘肽-S-转移酶活性的影响

用紫外分光光度法及荧光分光光度法测定中国对虾肝胰腺及鳃氨基比林-N-脱甲基酶(APND)、乙氧基香豆素-O-脱乙基酶(ECOD)、谷光甘肽-S-转移酶(GST)几种药物代谢酶的活性,探讨黄芩苷对上述几种酶活性的影响.实验结果表明,中国对虾肝胰腺和鳃丝APND活性分别为3.172和2.242 nmol/(min·mgprotein~(-1))、ECOD活性分别为0.325 0和0.357 9 pmol/(min·mgprotein-~(-1))、GST活性分别为70.59和13.97 nmol/(min·mgprotein~(-1)).中国对虾连续7 d投喂黄芩苷药饵(100 mg/kg)后,黄芩苷对几种酶均有不同程度的诱导作用,酶的活性均呈现先升高后降低的趋势.肝胰腺和鳃丝APND活性均在3 h左右达到最高,ECOD活性均在1 h左右达到最高,而GST活性迟于CYP450酶被诱导,2种组织GST活性分别在6,12 h达到最高水平.在临床用药中应考虑到黄芩苷对药物代谢酶的诱导作用,适当用药,合理配伍.