全氟辛烷磺酸对斑马鱼肝脏影响机制的代谢组学研究

为探究全氟辛烷磺酸(PFOS)对水生动物肝脏的慢性毒性毒害机制,寻找其肝脏慢性毒性相关的潜在标志物,运用气相色谱-质谱联用仪代谢组学方法研究暴露于PFOS中斑马鱼肝脏内源性代谢物的变化,寻找显著性差异代谢物及相关通路。实验分为4组:对照组及PFOS浓度分别为10、100、250μg·L~(-1)的实验组,每组20尾斑马鱼,实验时长40 d。结果显示,与对照组相比,PFOS浓度为10μg·L~(-1)的斑马鱼肝脏中筛选出4种发生显著性变化的代谢物,分别为牛磺酸、磷酸乙酰胺、β-D-葡萄糖、油酸,涉及4条代谢通路,即牛磺酸和亚牛磺酸代谢、鞘脂代谢、甘油磷脂代谢、糖酵解或葡萄糖生成。100μg·L~(-1)浓度组的斑马鱼肝脏中筛选出5种,包括牛磺酸、β-D-葡萄糖、棕榈酸、油酸、胆固醇,涉及5条代谢通路,即原代胆汁酸生物合成、牛磺酸和亚牛磺酸代谢、类固醇生物合成、类固醇激素生物合成、糖酵解或葡萄糖生成。250μg·L~(-1)浓度组的斑马鱼肝脏中筛选出5种,即牛磺酸、β-D-葡萄糖、乳酸、甘油-3-磷酸、磷酸乙酰胺,涉及5条代谢通路:牛磺酸和亚牛磺酸代谢、甘油磷脂代谢、甘油脂代谢、鞘脂代谢、糖酵解或葡萄糖生成。根据筛选出的差异代谢物生理功能和其涉及代谢通路分析,推测PFOS主要通过影响牛磺酸和亚牛磺酸代谢、糖酵解或葡萄糖生成及一些脂类代谢从而对斑马鱼肝脏产生毒性效应,其中牛磺酸、葡萄糖与油酸这3种代谢物可作为斑马鱼受PFOS胁迫肝脏代谢异常的潜在标记物。     

纳米TiO2对幼年小鼠血浆代谢谱影响的代谢组学研究

观察不同染毒剂量的纳米TiO2对ICR幼年小鼠血浆代谢谱变化的影响,寻找潜在的毒性效应的生物标志物.将幼年小鼠随机分为对照组以及10、30、100和300 mg·kg-1不同纳米TiO2染毒组进行28 d短期毒性试验,利用代谢组学技术初步筛选出TiO2毒性作用潜在的生物标志物,运用KEGG数据库对生物标志物进行拓扑分析;并利用靶向定量技术检测血浆胆酸、去氧胆酸、熊去氧胆酸和牛磺胆酸的含量变化,从而探究TiO2毒性作用相对应的生物标志物.从血浆中筛选出49种差异代谢物,发现纳米TiO2染毒后主要导致磷脂类及胆汁酸类化合物的异常.小鼠经过纳米TiO2 28 d染毒后,其毒性作用可能与磷脂代谢及胆汁酸代谢异常相关,血浆中的胆汁酸可以作为纳米TiO2产生肝脏毒性作用的生物标志物.     

啶虫脒对斑马鱼血浆代谢物的影响

为了辨析亚致死剂量啶虫脒对斑马鱼生长代谢的影响,采用气质联用技术,分析斑马血浆代谢物含量的变化.研究结果表明,亚致死剂量的啶虫脒诱导斑马鱼血浆多种代谢物含量上升,涉及到氨基酸代谢、糖代谢、三羧酸循环以及遗传物质合成多条代谢途径.亚致死剂量啶虫脒能扰乱非靶标生物的正常代谢,进而产生慢性毒性,所以亚致死剂量农药对非靶标生物的影响不可忽视.     

基于高分辨质谱研究双酚A对小鼠血浆代谢谱的影响

观察不同染毒剂量的双酚A(BPA)对C57BL6小鼠血浆代谢谱的影响,探讨其可能的毒性机制,并探寻毒性相关生物靶点。将小鼠随机分为对照组及1、10、50和250μg·kg-1组。利用高分辨质谱技术结合主成分分析、偏最小二乘分析等方法对血浆数据进行分析,发现对照组与各剂量组基本分离,并随染毒剂量的增加,各组逐渐偏离对照组,提示小鼠机体具有发生紊乱的趋势;运用Pathway Analysis数据库对差异性代谢物进行拓扑分析;运用Cytoscape和Metscape进行代谢产物的相关性分析和模块化分析。从血浆中筛选出27种差异代谢物,发现BPA的干预作用可能与亚油酸代谢、花生四烯酸代谢、丙氨酸、天冬氨酸和谷氨酸代谢、丙酮酸代谢、鞘脂类代谢和磷酸肌醇代谢等通路有关;对血浆差异性代谢产物的相关性分析和模块化分析结果表明,7个模块之间关系密切,BPA的干预作用可能与血浆7个模块代谢物的差别相关; BPA染毒14 d后,其毒性作用机制可能与丙酮酸、花生四烯酸、亮氨酸和亚油酸等生物靶点的异常相关,丙酮酸、花生四烯酸、亮氨酸和亚油酸等生物靶点可作为BPA引起机体毒性的标志物。     

代谢组学技术在环境毒理学研究中的应用

代谢组学作为系统生物学的一部分,通过考察机体受刺激后体液或组织中内源性代谢物的动态变化规律,并结合生物信息统计方法,可系统全面地揭示内因和外因作用于机体的毒性效应和机制。代谢组学技术具有快速、灵敏度高、选择性强的特点,逐渐在低剂量环境污染物长期暴露的毒性效应评估方面发挥出优势。本文综述了代谢组学技术的主要研究手段,在毒理学研究中的发展历程和优点,以及在环境毒理学研究中的应用及前景展望。重点讨论了代谢组学技术在重金属和持久性有机污染物(POPs)毒性评估以及环境胁迫耐受性评价中的应用。     

NMR代谢组学在水生软体动物生态毒理研究中的应用

随着核磁共振(NMR)技术的不断发展,基于NMR的代谢组学方法已经逐渐成为研究水生动物在代谢水平响应环境变化的一种有效方法。水生软体动物作为重要的环境监测生物,近年来人们在其相关研究中取得了不小的成果。文章总结了利用NMR的代谢组学方法研究各类环境胁迫因子对水生软体动物代谢影响的成果,主要综述了水生软体动物不同组织内代谢物组成和含量对环境胁迫的响应,同时对水生软体动物生态毒理代谢组学研究中存在的问题和今后的发展方向进行了讨论和展望。     

基于质谱的高覆盖代谢组学数据采集策略研究进展及在环境毒理学的应用

代谢组学是分析细胞或生物体中小分子化合物的一门组学科学.代谢组学的目标是尽可能分析特定生物样品中的所有代谢物,实现高覆盖分析.基于质谱的技术促进了组学规模的代谢物检测,成为代谢组学的重要分析平台.质谱数据采集策略的选择对获得全面且高质量的代谢谱图信息至关重要,影响数据采集率进而影响代谢组学的覆盖率.本文全面综述了当前基于质谱的高覆盖代谢组学数据采集策略的研究进展,总结方法的优势和局限,并概述当前代谢组学方法在环境毒理学领域的新应用,最后讨论当前局限并对未来发展做出展望.     

基于HepG2细胞脂质组学方法对黄浦江水质的综合评价

环境水体中复合污染的毒性识别一直是环境科学界关注的热点.基于代谢组学的生物监测方法通过监测暴露前后生物内源性代谢物的变化,在评估复合污染导致的综合毒性效应中具有广泛的应用前景.本研究从污染物对细胞脂类代谢的影响角度出发,采用体外肝癌细胞(HepG2)暴露和脂质组分析相结合的方法,对黄浦江干流和支流共27个采样点的水质进行了综合评价.研究发现,水样对HepG2细胞的脂代谢影响程度与采样点所属河段紧密相关.黄浦江中游水样对HepG2细胞的平均脂代谢影响程度(3.40％)高于上游(1.65％)和下游(0.78％),而流经城区的支流水样对细胞的脂代谢影响(5.20％)明显高于干流水样(1.80％).此外,在采集的水样中超过70％的样品都导致HepG2细胞内甘油三酯(TG)大量蓄积,揭示了黄浦江水体中的复合污染物可能诱导肝脏产生脂毒性损伤.本研究为环境水质监测提供新的方法,研究结果为上海市生态环境和水质安全管理部门提供重要的参考信息.     

十溴联苯醚（BDE209）在虹鳟体内的羟基代谢产物及其对甲状腺激素水平影响的初步研究（Preliminary Research on Hydroxylated Metabolites of Decabromodiphenyl Ether and Their Effects on Thyroid Hormone in Rainbow Trout（Oncorhynchus mykiss））

十溴联苯醚（Decabromodiphenyl ether, BDE209）被认为是可疑甲状腺素（TH）干扰物,但干扰机制目前尚不清楚. 采用单次腹腔注射的暴露方式,考察了在28d的实验周期内,不同浓度组的十溴联苯醚在虹鳟（Oncorhynchus mykiss）体内的羟基化代谢产物浓度与甲状腺激素（T3和T4）水平. 结果表明：十溴联苯醚在虹鳟的肝脏和血液中均可代谢为低溴代羟基多溴联苯醚,且羟基代谢产物在整个实验周期中浓度不断累积;与此同时,血浆中T3和T4浓度均出现下降趋势. 进一步的统计分析结果表明,羟基代谢产物与T3和T4浓度水平分别呈显著的负线性相关关系,这说明血浆中的甲状腺激素水平下降可能是由羟基化产物所引起的.     

十溴联苯醚(BDE209)在虹鳟体内的羟基代谢产物及其对甲状腺激素水平影响的初步研究

十溴联苯醚(Decabromodiphenyl ether,BDE209)被认为是可疑甲状腺素(TH)干扰物,但干扰机制目前尚不清楚.采用单次腹腔注射的暴露方式,考察了在28d的实验周期内,不同浓度组的十溴联苯醚在虹鳟(Oncorhynchus mykiss)体内的羟基化代谢产物浓度与甲状腺激素(T3和T4)水平.结果表明:十溴联苯醚在虹鳟的肝脏和血液中均可代谢为低溴代羟基多溴联苯醚,且羟基代谢产物在整个实验周期中浓度不断累积;与此同时,血浆中T3和T4浓度均出现下降趋势.进一步的统计分析结果表明,羟基代谢产物与T3和T4浓度水平分别呈显著的负线性相关关系,这说明血浆中的甲状腺激素水平下降可能是由羟基化产物所引起的.     

联合基于野外的代谢组学和化学分析手段筛选优先关注新兴污染物：在五大湖流域的案例研究

由于毒性评估项目很难与日渐增长的需要测试的污染物保持同步,所以较难将关注点集中在影响水生生态系统的最为生物相关的污染物上。由于评估潜在毒性污染物所造成的生物影响已被证明是有效的,内生性代谢物的研究（代谢组学）对于剔除那些较低可能造成生物影响的污染物或许有一定帮助,从而找出生物重要性最高的污染物。本研究在北美五大湖流域的18个地点针对置于笼中的黑头软口鲦（Pimephales promelas）进行实验。我们测定了水体温度和水样中的污染物浓度（目标污染物132种,检出86种）,并使用1H-NMR谱测量了肝极性提取物中的内生性代谢物。利用偏最小二乘法回归来比对内生性代谢物的相对丰度与污染物浓度和环境温度。结果表明内生性极性代谢物的指标与最多49种污染物存在共同变化。因此我们认为至多52%的检出污染物与内生性代谢物变化的共同变化不显著,表明这些污染物很可能不会在这些地点造成可以检测到的影响。这是通过缩短对于实验地点有着潜在影响的污染物列表从而扫描出检出污染物生物相关性的第一步。类似的信息有助于风险评估者区分不同污染物的重要性并将重点毒性测试放在最为生物相关的污染物上。
精选自Nicol Janecko, Lucie Pokludova, Jana Blahova, Zdenka Svobodova, Ivan Literak. Linking field-based metabolomics and chemical analyses to prioritize contaminants of emerging concern in the Great Lakes basin. Environmental Toxicology and Chemistry: Volume 35, Issue 10, pages 2493–2502, October 2016. DOI: 10.1002/etc.3409
详情请见http://onlinelibrary.wiley.com/doi/10.1002/etc.3409/full
     

Water-dispersible nano-pollutions reshape microbial metabolism in type-specific manners: A metabolic and bacteriological investigation in Escherichia coli

• Water-dispersible nano-pollutions exhibit type-specific toxic effects on E. coli. • Global metabolite profiling was used to characterize metabolic disruption patterns. • Key dysregulated metabolites responsive to nano-pollution exposures were found. • Amino acid metabolism and purine metabolism are perturbed at nano-pollutions. Incomplete separation and recycling of nanoparticles are causing undesirable nanopollution and thus raising great concerns with regard to nanosafety. Since microorganisms are important regulator of physiological processes in many organisms, the interaction between nanopollution and microbial metabolomics and the resultant impact on the host’s health are important but unclear. To investigate how typical nanopollution perturbs microbial growth and metabolism, Escherichia coli (E. coli) in vitro was treated with six water-dispersible nanomaterials (nanoplastic, nanosilver, nano-TiO₂, nano-ZnO, semiconductor quantum dots (QDs), carbon dots (CDs)) at human-/environment-relevant concentration levels. The nanomaterials exhibited type-specific toxic effects on E. coli growth. Global metabolite profiling was used to characterize metabolic disruption patterns in the model microorganism exposed to different nanopollutants. The percentage of significant metabolites (p<0.05, VIP>1) accounted for 6%–38% of the total 293 identified metabolites in each of the nanomaterial-contaminated bacterial groups. Metabolic results also exhibited significant differences between different nanopollutants and dose levels, revealing type-specific and untypical concentration-dependent metabolic responses. Key metabolites responsive to nanopollution exposures were mainly involved in amino acid and purine metabolisms, where 5, 4, and 7 significant metabolic features were included in arginine and proline metabolism, phenylalanine metabolism, and purine metabolism, respectively. In conclusion, this study horizontally compared and demonstrated how typical nanopollution perturbs microbial growth and metabolomics in a type-specific manner, which broadens our understanding of the ecotoxicity of nanopollutants on microorganisms.     

Metabolic responses in zebrafish (Danio rerio) exposed to zinc and cadmium by nuclear magnetic resonance -based metabolomics

Heavy metals are common marine and soil pollutants that are mainly the result of industrial activity, and are a threat to the environment and human health. In this study, ¹H nuclear magnetic resonance (NMR)-based metabolomics was applied to adult Danio rerio to monitor the metabolic change as a response to ZnCl₂ and CdCl₂ exposure at different concentrations for 72?h. NMR spectroscopy was used to identify and quantify the metabolites extracted from D. rerio. The metabolite profiles of the control and heavy metal exposed group were classified by partial least squares – discriminant analysis (PLS-DA) analysis, and potential contaminant-specific biomarkers were suggested. For the ZnCl₂-exposed zebrafish, the levels of ATP, aspartate and NAD⁺ were increased, whereas the levels of formate, inosine, hypoxanthine and succinate decreased. In addition, the CdCl₂-exposed zebrafish showed an increase in the levels of ATP and formate and a decrease in the levels of glutamate, inosine and glutathione. Overall, Zn and Cd may lead to neurotoxicity, disturbances in the energy metabolism and oxidative stress. Our finding demonstrated that the application of NMR-based metabolomics might be useful for detecting the toxicity caused by sub-lethal concentrations of heavy metal contaminants in the environment.     

核受体超家族及其酵母双杂交检测技术

环境污染物的内分泌干扰问题近年来引起了极大的关注,大量研究证实:核受体是内分泌干扰的重要作用位点.论文在总结国内外相关研究基础上,对生物体内核受体超家族的组成、结构特征、作用模式进行了概括总结;对环境内分泌干扰物干扰核受体超家族最新研究进展给予了评述;对酵母双杂交检测技术的原理及其在核受体超家族和环境内分泌干扰效应研究中的应用进行了探讨,指出应将酵母双杂交检测技术与核受体超家族研究相结合,建立成组重组核受体基因双杂交酵母体系,应用于多种内分泌干扰效应和作用机制的系统研究.     

Interactions of Endocrine-active environmental chemicals with the nuclear receptor PXR

There is growing concern about the human-health impact of environmental chemicals that have the potential to disrupt normal endocrine function. Endocrine disrupting chemicals (EDCs) include structurally diverse organochlorine pesticides, polychlorinated biphenyls (PCBs), plasticizers, fungicides, herbicides and pharmaceutical compounds, and can have a profound impact on development, and on reproductive, neurological and immune system functions. While many studies have focused on the role of androgen receptor, estrogen receptor and aryl hydrocarbon receptor in mediating the effects of EDCs, other nuclear receptors that regulate steroid hormone action and metabolism may also serve as targets of EDC action. This review focuses on two classes of EDCs, PCBs and phthalate monoesters, both of which have been shown to interact with pregnane X receptor (PXR), a member of the nuclear receptor superfamily that regulates a large number of target genes, many of which have important roles in steroid metabolism and transport. Recent findings on the ability of PCBs and phthalate monoesters to activate PXR are discussed and the potential role of PXR and other intracellular receptor proteins in mediating toxicities associated with EDC exposure is considered. Finally, we discuss several gaps in our knowledge regarding the actions of EDCs and the difficulties associated with the evaluation of risks associated with exposure to these endocrine active environmental chemicals.     

氯吡嘧磺隆和镉复合对斑马鱼肝脏的影响

为了探究污染物镉和氯吡嘧磺隆复合污染对非靶标生物斑马鱼的毒性作用,通过急性毒性试验,获得镉和氯吡嘧磺隆对斑马鱼的LC₅₀。以LC₂₅作为0水平,设置9个处理组进行试验,检测不同处理组斑马鱼96 h死亡率、斑马鱼肝脏中CAT、MDA和羧酸酯酶CarE。检测结果表明,镉和氯吡嘧磺隆对斑马鱼存在交互作用,当水体中镉浓度为3.12 mg·L^-1,氯吡嘧磺隆浓度为14.03 mg·L^-1,两种物质复合对斑马鱼的毒性效应最大。将毒性效应最大处理组的斑马鱼肝脏代谢物进行采样分析,研究结果表明,镉和氯吡嘧磺隆复合造成斑马鱼肝脏核苷酸、乙醛酸、能量、氨基酸等代谢途径异常,多种代谢物含量显著下降。研究结果可为水环境中氯吡嘧磺隆和镉复合污染的生态风险评价和早期预警提供依据。     

受体报告基因实验及其在维甲酸和维甲酸X受体干扰物监测中的应用

受体报告基因实验具有快速、经济、灵敏、方便等诸多优势,在高通量筛选类或抗雌雄激素等通过核受体起作用的环境内分泌干扰物方面得到了广泛的应用。环境中的维甲酸和维甲酸X受体干扰物如有机锡等有着类似的作用机制,研究者也开始采用受体报告基因实验的方法对该类污染物进行筛选与监测。本文综述了受体报告基因实验的技术方法,包括报告基因和宿主细胞的选择,并介绍了该方法在人工合成的维甲酸和维甲酸X受体干扰物筛选以及环境样品中该类污染监测中的应用。综述总结了应用受体报告基因实验检测环境内分泌干扰物研究中的不足并对该方法的未来发展进行了展望,希望为该方法在环境监测和评估中的应用提供新的思路。     

Untargeted metabolomic analysis of pregnant women exposure to perfluorooctanoic acid at different degrees

● Metabolome can distinguish pregnant women exposure to PFOA at different degrees. ● Metabolome can reveal the metabolic changes of pregnant women exposure to PFOA. ● PFOA exposure degrees could affect the GSH metabolism of pregnant women. ● PFOA exposure degrees could change the microbiota metabolism of pregnant women. Perfluorooctanoic acid (PFOA) is a novel type of persistent synthetic organic pollutant, and its exposure on pregnant women can cause some adverse effects, such as pregnancy-induced hypertension, gestational diabetes mellitus, and preeclampsia. Therefore, understanding the metabolic changes caused by PFOA exposure is of great significance to protect pregnant women from its adverse effects. In this study, the metabolomes from the urine samples of pregnant women exposure to PFOA at different degrees were analyzed by GC-MS and LC-MS. The samples in different groups were distinguished and the differential metabolites were screened based on the VIP value, FC, and P-value of each comparison group through multivariate statistical analysis. The pathways related to differential metabolites were searched to reveal the effects of PFOA exposure on metabolic changes in pregnant women at different degrees. Finally, the ROC of differential metabolites was performed, and the differential metabolites with large area under the curve (AUC) values were selected and compared to identify the mutually differential metabolites. Meanwhile, these metabolites were fitted with a multivariable to explore if they could be used to distinguish different groups. The quantitative comparison of mutually differential metabolites revealed that the levels of L-cysteine, glycine, and 5-aminovaleric acid were positively correlated with the degree of PFOA exposure, indicating that different degrees of PFOA exposure could affect the synthesis or degradation of GSH and change the metabolism of oral or intestinal microbiota. Additionally, they may cause oxidative stress and abnormal fat metabolism in pregnant women.