Comparision of the waterborne and dietary routes of exposure on the effects of Benzo(a)pyrene on biotransformation pathways in Nile tilapia (Oreochromis niloticus)

BaP is one of the most studied PAH, due to its ubiquitous presence in aquatic environments and toxicity to aquatic organisms. The main goal of this study was to assess BaP effects in Nile Tilapia after waterborne and dietary exposures, through the evaluation of EROD and GST activities in liver, gills and intestine, and BaP metabolites in bile; and also to evaluate the usefulness of these commonly used biomarkers after two different routes of exposure. Waterborne exposure to BaP led to a significant induction of EROD in all tissues analyzed (644%, 1640% and 2880% in relation to solvent in liver, gill and intestine respectively) while in dietary exposures EROD was induced only in intestine (3143%) after exposure to high BaP concentrations. GST activities with CDNB were slightly induced in liver (40%) and in gill (66%) after water exposure to BaP, and in intestine after dietary exposure to low BaP concentrations (182%). BaP metabolites in bile increased after both exposure routes, and were highly correlated with EROD activity after water exposure. In summary, this work has shown that the effects of BaP on biotransformation pathways depend on the route of exposure. Moreover, barrier tissues like gills and intestine also have an important role in the first-pass metabolism of BaP, reducing the amount of parent compound that reaches the liver to be metabolized. For that reason, EROD activity as a biomarker of exposure should also be applied in extrahepatic organs, like gills and intestine, in monitoring studies. Biliary BaP type metabolites are good reflectors of contamination levels under both exposure routes, while GST activity with CDNB as substrate, as a phase II enzyme, does not seem a reliable biomarker of exposure to BaP regardless the route of exposure.     

苯并芘对稀有鮈鲫的内分泌干扰效应研究

为评价典型有机污染物对长江三峡库区上游特有珍稀鱼类的环境风险,本实验以稀有鮈鲫为对象,研究了低剂量多环芳烃(PAHs)暴露而引起的内分泌干扰效应.将成年稀有鮈鲫暴露于不同浓度(0、0.1、0.3、1.0和3.0μg·L-1)的苯并芘(Ba P)中28 d,测定了体质系数、性激素含量、下丘脑-垂体-性腺(HPG)轴和肝脏中与繁殖相关基因的表达.结果显示:在雌鱼中,Ba P暴露后,睾酮(T)的含量显著升高,雌二醇(E2)的水平显著下降并改变了T/E2的比值;在雄鱼中,T和E2的含量都显著下降.雌雄鱼性激素的改变可能与类固醇合成相关基因(Cyp17、Cyp19a)表达量的改变有关.Ba P暴露显著抑制了雌雄鱼肝脏中卵黄蛋白原(VTG)基因的表达.以上检测到的激素或者基因表达的改变主要发生在高剂量(3.0μg·L-1)Ba P暴露下,而长江上游库区水体中多环芳烃含量较低(100 ng·L-1),因此推测,库区存在的多环芳烃不会对三峡库区上游水体中的鱼类产生内分泌干扰效应.     

Chemopreventive effects of ellagic acid against genotoxicity induced by benzo(a)pyrene

Benzo(a)pyrene (B(a)P) is a commonly used indicator for polycyclic aromatic hydrocarbons (PAHs) contamination. The use of certain phytochemicals and some polyphenolic compounds proved to be effective in blocking PAH-induced effects. The aim of the present study was to examine the possible inhibitory effects of ellagic acid (EA), present in berries and nuts, against B(a)P-mediated toxicity using in vitro and in vivo test systems. In vitro method included the Ames test, using Salmonella typhimurium tester strain TA100. In vivo experiments were based on mouse bone marrow micronucleus (MN) assay and immunoblotting. EA produced a decrease in the number of revertant colonies against B(a)P in the Ames assay. Further, there was a reduction in the mean number of micronucleated polychromatic erythrocytes in the presence of EA in the co-treatment protocol of the MN test. Western blotting results showed downregulation of CYP450 isoform CYP1A1 which prevented bioactivation of B(a)P. Thus, the present study demonstrates antimutagenic role played by EA against the mutagenic activity of B(a)P in both in vitro and in vivo systems.     

基于贝叶斯的苯并(α)芘PBPK模型优化与健康风险评估应用

应用基于生理的药代动力学(PBPK)模型预测苯并(α)芘(BaP)暴露的人体内部剂量,基于贝叶斯的马尔科夫链蒙特卡洛模拟(MCMC)方法对模型参数进行校准和优化,最后运用已优化的模型对BaP内暴露基准值进行推导.研究发现,基于贝叶斯的MCMC方法对模型后验参数校准后,模型精度明显提高,两个数据集验证结果显示残差平方和分别降低了72%和94%.PBPK模型以BaP和子代谢物3-羟基苯并(α)芘(3-OHBaP)的体内动力学过程为结构基础,模拟BaP体内浓度分布大小为脂肪>肾脏>皮肤>缓慢灌注组织>快速灌注组织>静脉血>肝脏;3-OHBaP体内浓度分布大小为肾脏>快速灌注组织>脂肪>肺>静脉血>缓慢灌注组织>肝脏>皮肤.敏感性分析显示,快速灌注组织-血分配系数对模型输出影响最大,灵敏度系数超过了200%;排泄系数影响最小,只有肾小球过滤率K_BR的灵敏度系数超过了1%.以美国国家环境保护局推荐的参考浓度2.0×10^-6mg/m³为外暴露安全基准值,基于PBPK模型推导了职业暴露的BaP生物监测当量(BE),结果显示BE值为0.405pmol/mol肌酐(尿液3-OHBaP平均浓度),为基于人体内暴露剂量水平进行定量健康风险评估奠定了基础.     

Modeling the exposure of children and adults via diet to chemicals in the environment with crop-specific models

Exposure to chemicals via diet is a major uptake pathway for many compounds but is often estimated in a rather generic way. We use a new model framework (NMF) with crop-specific models to predict the dietary intake by 4-5-year-old children and 14-75-year-old women of three environmental compounds from their background concentrations in soil and air. Calculated daily intakes of benzo(a)pyrene and 2,3,7,8-TCDD are in good agreement with measured results from diet studies. The major source of both compounds in human diet is deposition from air. Inhalation of air and ingestion of soil play a minor role. Children take up more than twice the amount than adults per kg bodyweight, due to higher consumption per kg bodyweight. Contrary, the methods for indirect human exposure suggested in the Technical Guidance Document (TGD) for chemical risk assessment in the EU lead to overprediction, due to unrealistic consumption data and a false root model.     

兰州市大气中苯并(a)芘与儿童尿中1-羟基芘的分析及风险评价

对兰州市不同区域空气中苯并(a)芘和儿童尿液中1-羟基芘同步采样,分别采用气相色谱-质谱联用法和高效液相色谱法进行检测.结果表明:不同区域苯并(a)芘浓度有显著差别,研究区日均浓度约为对照区日均浓度的两倍.冬季大气中的浓度明显高于夏季,其中研究区冬季均值约为夏季的26.46倍,对照区冬季均值约为夏季的29.41倍.儿童尿中1-羟基芘的浓度与空气中苯并(a)芘的浓度呈一致的变化,冬季研究区儿童尿中1-羟基芘的均值为0.86μmol·(mol肌酐)-1,对照区为0.63μmol·(mol肌酐)-1.冬季研究区儿童的终生超额危险度最高,达到2.32×10-4,接近不可接受水平.     

蜈蚣草中芘与砷的赋存特征及其相互作用

超富集植物蜈蚣草能够累积土壤环境中的砷,为探明蜈蚣草植株中芘的赋存特征及其与砷的相互作用,采用双光子激光共聚焦扫描显微技术检测观察芘在该植物中赋存和分布规律.结果表明,添加芘导致蜈蚣草各部位砷含量显著降低,其中叶部和茎部砷含量下降幅度达35%左右,根部砷含量减少20.5%.芘的添加使得植物各部位三价砷比例下降,五价砷比例升高,蜈蚣草叶部三价砷和五价砷赋存浓度下降幅度最大,下降幅度分别为42.2%和32.49%.添加砷可促进蜈蚣草根部和茎部中芘的积累,每株分别增加了9.8μg和139 ng,但对叶部中芘的积累无显著影响.芘主要分布于蜈蚣草细胞膜与细胞内部的细胞器膜结构及细胞核核膜附近,但细胞质中芘的含量较少,茎部韧皮部与皮层以及叶部栅栏组织和海绵组织中几乎没有检测到芘.     

采用固定化技术处理土壤中菲、芘污染物

采用动胶杆菌(Zoogloea sp.)固定化技术包埋来降解土壤中菲、芘污染物,在不同接种量、不同系列浓度下,对降解效果进行了测定.结果表明:5%的接种量最为合理,在168h时,固定化细菌对菲、芘的降解率达84.89%和76.94%;而在相同条件下土著菌降解仅达到27.85%和19.65%,因此动胶杆菌(Zoogloea sp.)具有更好的降解能力.另外,还用电镜观察研究了动胶杆菌在载体中的分布形态,表明了固定化细菌降解土壤中菲、芘具有较高的优势.     

用连续提取法研究芘在鱼鳃表面的结合形态

建立了研究鱼鳃表面芘的结合状态的四步连续提取方法．分别用H2O、CaCl2、CH3OH和n-C6H14+CH2Cl2(1:1, v/v)混合液提取经暴露的鱼鳃．操作定义的前3步分别提取松散附着在鳃表的鱼鳃黏液中的芘,紧密附着在鳃表的鱼鳃黏液中的芘以及直接吸着在鱼鳃表皮上的芘,第4步则用来提取鱼鳃组织吸收的芘．其提取的芘分别占总量的 4．0％、12．1％、45．2％和38．7％．经3 h暴露,进入鳃组织的芘仅为总测定量的三分之一强,因此大部分芘并未进入鳃组织．单独进行的五步CH3OH连续提取实验结果证明,CH3OH提取步骤能有效地解吸鳃表吸着的芘,但不会导致鳃组织中芘的释放．动态实验结果表明,黏液结合态芘在暴露1h左右即达到平衡,而鳃表吸着态和鳃组织吸收态芘在暴露3h内含量迅速上升,3 h后显著趋缓．     

Reduced-glutathione concentrations in Boleophthalmus pectinirostris tissues exposed to benzo(a)pyrene

ＩｎｔｒｏｄｕｃｔｉｏｎＢｅｉｎｇａｔｙｐｉｃａｌｐｏｌｙｃｙｃｌｉｃａｒｏｍａｔｉｃｈｙｄｒｏｃａｒｂｏｎｓ(ＰＡＨｓ) ,ｂｅｎｚｏ[ａ]ｐｙｒｅｎｅ(ＢａＰ)ｄｅｓｅｒｖｅｓｓｅｒｉｏｕｓｓｔｕｄｙｂｅｃａｕｓｅＰＡＨｓａｒｅｎｏｗｒｅｃｏｇｎｉｚｅｄａｓｍａｊｏｒｅｎｖｉｒｏｎｍｅｎｔａｌｐｏｌｌｕｔａｎｔｓ .ＥｘｐｏｓｕｒｅｔｏＰａＨｓｉｓｖｉｒｔｕａｌｌｙｕｎａｖｏｉｄａｂｌｅａｎｄｉｓｓｔｒｏｎｇｌｙｓｕｓｐｅｎｄｅｄｏｆｂｅｉｎｇａｃａｕｓａｔｉｖｅｆａｃｔｏｒｉｎｃｅｒｔａｉｎｔｕｍｏ…