Genome-wide identification and characterization of cytochrome P450 monooxygenase genes in the ciliate Tetrahymena thermophila

Background: Cytochrome P450 monooxygenases play key roles in the metabolism of a wide variety of substrates and they are closely associated with endocellular physiological processes or detoxification metabolism under environmental exposure. To date, however, none has been systematically characterized in the phylum Ciliophora. T. thermophila possess many advantages as a eukaryotic model organism and it exhibits rapid and sensitive responses to xenobiotics, making it an ideal model system to study the evolutionary and functional diversity of the P450 monooxygenase gene family. Results: A total of 44 putative functional cytochrome P450 genes were identified and could be classified into 13 families and 21 sub-families according to standard nomenclature. The characteristics of both the conserved intron-exon organization and scaffold localization of tandem repeats within each P450 family clade suggested that the enlargement of T. thermophila P450 families probably resulted from recent separate small duplication events. Gene expression patterns of all T. thermophila P450s during three important cell physiological stages (vegetative growth, starvation and conjugation) were analyzed based on EST and microarray data, and three main categories of expression patterns were postulated. Evolutionary analysis including codon usage preference, sit-especific selection and gene-expression evolution patterns were investigated and the results indicated remarkable divergences among the T. thermophila P450 genes. Conclusion: The characterization, expression and evolutionary analysis of T. thermophila P450 monooxygenase genes in the current study provides useful information for understanding the characteristics and diversities of the P450 genes in the Ciliophora, and provides the baseline for functional analyses of individual P450 isoforms in this model ciliate species.

植物细胞色素P450对土壤PAHs污染早期诊断研究

以高等植物为试验材料，建立了细胞色素P450含量的测定方法。在此基础上通过对5种植物（小麦、玉米、白菜、萝卜、大豆）的筛选试验，最终选择小麦绿苗和玉米黄化苗作为供试材料，进行土壤菲、芘单一与复合污染条件下，细胞色素P450含量与多环芳烃污染的量-效关系生态毒理诊断研究。同时为比较不同指标对污染物生态毒性敏感性，进行了相同剂量条件下，SOD、POD、CAT活性,高等植物种子发芽与根伸长抑制率与菲、芘单一与复合污染诱导的剂量-效应关系试验研究。 细胞色素P450含量污染诱导研究结果表明，在较低的浓度下，土壤菲、芘单一暴露与小麦和玉米幼苗P450含量具有显著的污染诱导剂量-效应关系，菲、芘复合污染对小麦和玉米的P450产生协同毒性作用。两种植物P450含量对菲、芘污染指示较为敏感。 SOD、POD、CAT活性比较试验结果表明，SOD、POD、CAT活性对菲、芘污染具有响应，但响应敏感性不及植物色素P450，有些不能进行定量诊断，但可用于定性诊断。 发芽率和根伸长抑制率试验比较结果表明，在同样的供试浓度范围内，小麦发芽率和根伸长与对照相当，未出现明显诱导或抑制的响应。植物色素P450含量对菲、芘单一与复合污染的指示作用比植物种子发芽率和根伸长抑制率更为敏感。 植物细胞色素P450这一敏感指标可以作为生物标记物，与化学分析及抗氧化酶系指标结合起来进行土壤PAHs早期诊断。既有助于提高指标的准确性和敏感性，推动形成完善的土壤污染生态毒理诊断方法，还可从生理和分子水平使污染诱发障碍的机理等得到破译。

Structural determinants of steroids for cytochrome P450 3A4-mediated metabolism

Cytochrome P450 3A4 (CYP3A4), a major member of cytochrome P450 isoenzymes, metabolizes the majority of steroids in 6beta-position. For the purpose of determining requisite structural features of a series of structurally related steroids for CYP3A4-mediated metabolism, three-dimensional pharmacophore modeling as well as electrotopological state map were conducted for 15 steroids. Though prior studies speculated that the chemical reactivity of the allylic 6beta-position might have a greater influence on CYP3A4 selective 6-hydroxylation than steric constraints in the enzyme, our results reveal that for CYP3A4 steroidal substrates, it is not the chemical reactivity of atoms at 6beta-site, but the pharmacophoric features, i.e. the two hydrophobic rings together with two H-bond donors, that act as the key factors responsible for detemining the CYP3A4 selective 6-hydroxylation of steroids. (C) 2004 Elsevier B.V. All rights reserved.

蚯蚓(Eiseniafetida)细胞色素P450及抗氧化酶系对环境浓度苯并(a)芘的响应

以赤子爱胜蚓(Eisenia fetida)为供试生物,草甸棕壤为供试土壤,以蚯蚓微粒体细胞色素P450含量、抗氧化酶系以及谷胱甘肽转移酶活性为指标,进行了土壤中苯并(a)芘[B(a)P]暴露与酶活性的量-效关系研究。结果表明,在接近沈抚灌区实地污染状况B(a)P(0.1～2.0mg.kg-1)暴露下,第1、3、7以及第14d取样时,P450和SOD有较好的响应:SOD在第1、3d显著升高,而在第7、14d时降低;P450总体表现为低浓度B(a)P诱导下降低,高浓度下升高的趋势。CAT、POD以及GST的敏感性相对较差。比照其他4个指标,蚯蚓P450的敏感性更为优异,具有较好的应用前景。指标敏感性总体表现为:P450>SOD>CAT/POD>GST。

真菌细胞色素P450与多环芳烃浓度及降解率的相互关系

以菲、芘作为多环芳烃代表污染物,采用室内培养方法,研究了4种真菌细胞色素P450含量与多环芳烃(PAHs)浓度及降解率的相互关系。结果表明,在一定的浓度范围内(菲0～200mg·L-1、芘0～100mg·L-1),菲、芘浓度与真菌P450含量呈明显正相关,菲、芘浓度与真菌P450含量之间表现出明显的剂量-效应关系。在上述浓度内,真菌降解多环芳烃的能力与真菌P450含量之间也表现出明显的剂量-效应关系。在选用的4株真菌中,Zj1(镰刀菌)降解菲和芘的能力最强,Zj3(小克银汉)次之,Zj2(毛霉)和Zj4(青霉)降解能力较弱。

细胞色素P—450模拟体系的活性中间体-氧配位铬(V)四苯基卟啉配合物的分离、表征和氧化反应

氮气保护下二氯甲烷中铬(Ⅲ)四苯基卟琳衍生物在-40℃与亚碘酰苯反应,分离得氧配位铬(V)四苯基叶啉配合物:O=Cr(V)TPP(CI)PhI,O—Cr(V)TPP(N_3)PhI,O—Cr(V)TPP(p-CH_3O-C_6H_4O)1/2PhI.已经元素分析、可见、红外、顺磁、核磁和质谱法结构表征.这些配合物能氧化苯乙烯,环己醇,环己烯和环己烷,可作为细胞色素P—450模拟体系的活性中间体.

鱼肝EROD酶活力诱导作为二噁的水生态毒理学指标

利用离体EROD法对严家湖各氧化塘鱼肝中二噁的毒性效应进行了定量测定，同时与HRGC／HRMS－MID化学分析结果和野外活体暴露鱼肝中EROD活力诱导结果进行了比较。研究发现，各氧化塘中鱼肝的离体、活体EROD测定结果与化学分析结果之间有着极好的相关性。这不仅揭示了鱼肝细胞色素P450系统以EROD酶活力诱导指示可作为二的水生态毒理学指标的可靠性和准确性，同时也表明了活体EROD和离体EROD生物监测方法两者可以相互补充，用于其他环境生物样品中二类毒物的快速筛选。

Waterborne exposure to fluorotelomer alcohol 6:2 FTOH alters plasma sex hormone and gene transcription in the hypothalamic-pituitary-gonadal (HPG) axis of zebrafish

Fluorotelomer alcohols (FTOHs) have shown estrogenic activity in vitro and in vivo, but the mechanism of this activity is not known. In this study, 18-week-old zebrafish (Danio rerio) were exposed to 0, 0.03, 0.3 and 3.0 mg/l 1H, 1H, 2H, 2H-perfluorooctan-1-ol (6:2 ETCH) for 7 days, and the effects on plasma sex hormone levels were measured followed by use of real-time PCR to examine selected gene expression in hypothalamic-pituitary-gonadal (HPG) axis and liver. Exposure to 6:2 FTOH significantly increased plasma estradiol (E2) and testosterone (T) levels in both males and females. Furthermore, the ratio of T/E2 was reduced in females while increased in males. In females, the increase of E2 was accompanied by up-regulated hepatic estrogenic receptor alpha (ER alpha) and vitellogenin (VTG1 and VTG3) expression. In males, the elevation of the T level is consistent with the up-regulation of cytochrome P450 c17 alpha-hydroxylase, 17, 20-lase (CYP17) and the down-regulation of cytochrome P450 aromatase A (CYP19A). The present study demonstrated that waterborne exposure to 6:2 FTOH alter plasma sex hormone levels and the ratio of T/E2, as well as the transcriptional profiles of some genes in the HPG axis and liver. The results suggested that FTOHs may disturb fish reproduction through endocrine disrupted activity. (C) 2009 Elsevier B.V. All rights reserved.