Investigating the hepatic toxicity of 3,3â-dichlorobiphenyl (PCB-11) in zebrafish (Danio rerio)

研究 3,3-二氯联苯 (PCB-11) 对斑马鱼 (Danio rerio) 的肝毒性

• 批准号: 10058206
• 负责人: Monika Roy
• 金额: $ 3.21万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2021-06-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10058206
• 关键词: Acute; Adolescent; Affect; Agonist; Aryl Hydrocarbon Receptor; Benzo(a)pyrene; Biochemistry; Biological Assay; Biological Sciences; CYP11A1 gene; Cell Culture Techniques; Cells; Child; Chronic; Cytochromes; Data; Deformity; Deposition; Development; Disease Progression; Dose; Economic Burden; Embryo; Environment; Environmental Pollution; Equipment; Etiology; Exposure to; Faculty; Fatty Acids; Fertilization; Fishes; Fluorescence Microscopy; Fluorescence-Activated Cell Sorting; Funding; Genes; Goals; Government; Growth; Growth and Development function; Half-Life; Hepatic; Hepatocyte; Hepatotoxicity; Histopathology; Hour; Human; Individual; Infant; Institutes; Institution; Learning; Life; Lipids; Liver; Liver diseases; Measures; Mentors; Mentorship; Methods; Microscopy; Modeling; Molecular; Molecular Biology; Morphology; Obesity; Oils; Organ; Outcome; Parents; Pathway interactions; Pharmacologic Substance; Phenols; Phenotype; Pigments; Polychlorinated Biphenyls; Population; Postdoctoral Fellow; Pregnant Women; Process; Public Health; Research; Resources; Rodent; Sampling; Scientist; Serum; Stains; Sulfate; Techniques; Time; Tissues; Toxic effect; Toxicity Tests; Toxicology; Training; Transgenic Organisms; United States National Institutes of Health; Universities; Vacuole; Work; Zebrafish; career preparation; enzyme activity; experience; in vivo; lipid metabolism; liver development; longitudinal analysis; member; non-alcoholic fatty liver disease; prevent; transcriptome sequencing; translational impact; urban area

! PROJECT SUMMARY/ABSTRACT Non-alcoholic fatty liver disease (NAFLD) affects 30-40% of the U.S. population and has an estimated U.S. economic burden of $103 billion per year. NAFLD is also prevalent in children and adolescents, affecting 34.2% of obese individuals and 7.6% of this population overall. Higher-chlorinated polychlorinated biphenyls (PCBs) are implicated in the etiology of NAFLD, however, little is known about the hepatic toxicity potential of lower-chlorinated PCBs, including 3,3’-dichlorobiphenyl (PCB-11), widely detected in environmental and human samples, including in pregnant women. Preliminary research shows that acute developmental exposures to PCB-11 alone do not result in overt toxicological outcomes in zebrafish (Danio rerio), however, it misregulates hepatic-associated genes, stunts liver development, and increases vacuolization in liver tissue. Further, in combination with Aryl hydrocarbon receptor (Ahr) agonists, PCB-11 can either inhibit Cytochrome p4501a (Cyp1a) enzyme activity induced by a model PAH, exacerbating toxicity, or prevent toxicity induced by PCB-126, highlighting the importance of using a mixtures approach. The objective of this proposed study is to investigate the hepatic toxicity of PCB-11 metabolites, which have longer half-lives in vivo for rodents, and in preliminary data of low-dose exposures show increased hepatic lipid deposition in 15-day fish. The central hypothesis of this proposed study is that PCB-11 induced hepatic toxicity is attributed to its metabolites and is associated with a NAFLD liver phenotype during juvenile development. Aim 1 will determine the toxicity contribution of phenolic and sulfate PCB-11 metabolites in 4-day old fish. Aim 2 will assess PCB-11 and PCB-11-Sulfate toxicity in 30-day juvenile stage fish after low-dose chronic single and mixture exposures. The proposed training plan to execute these aims includes learning and/or performing methods such as EROD assays to measure Cyp1a activity, fluorescence microscopy to assess liver development, histopathology, Oil- Red-O staining to assess lipid deposition, and Fluorescence-Activated Cell Sorting (FACS) for fatty acid composition analysis of hepatocytes. In addition, chronic exposures will be carried out using the OECD Fish, Early-Life Stage (FELS) toxicity test (OECD TG 210) to gain practical experience in regulatory toxicity approaches and to increase preparation for the career goal of working as a government scientist to assess the public health impacts of contaminant exposures. This project will yield in vivo mechanistic toxicity data on PCB-11 and its metabolites at several developmental stages to be of use for public health regulators. The sponsor, Dr. Alicia Timme-Laragy, has a record of NIH funding, is a recent mentor of an NIH F32 Postdoctoral Fellow (now a faculty member at San Diego State University), has numerous equipment for fluorescence microscopy, molecular biology and biochemistry work, and is capable of providing the mentorship and guidance needed to accomplish the goals of the proposed project. Further, UMass Amherst is a large R1 research institution and houses several resources, including the Institute for Applied Life Sciences (IALS). !

呢 项目摘要/摘要 非酒精性脂肪肝疾病（NAFLD）影响美国人群的30-40％，并估计美国 珀斯的经济负担为10030亿美元。 34.2％的肥胖个体和该人口的7.6％的总体氯化双苯基 （PCB）与NAFLD的病因有关，但是，对肝脏的肝脏势势知之甚少 氯苯基含有下氯的PCB，含有3,3'-二氯苯基（PCB-11），在象牙和 人类样本，包括孕妇 仅对PCB-11的暴露不会导致斑马鱼（Danio Rerio）的结果 置于肝相关的基因，特技肝发育并增加肝组织中的空置化。 此外，与芳基烃受体（AHR）激动剂结合使用，PCB-11可以eithibibibibibibitchrome 由模型PAH诱导的P4501A（CYP1A）酶活性，加剧毒性或预防由 PCB-126，突出了使用混合物方法的重要性。 研究PCB-11代谢产物的肝毒性，啮齿动物的体内半衰期更长，在 低剂量暴露的初步数据显示，中央的肝脂质沉积物增加。 支撑研究的假设是PCB-11诱导毒性归因于IS代谢物 在少年发育过程中与NAFLD肝脏表型相关。 酚类和硫酸盐PCB-11代谢物的毒性贡献为4天的AIM 2。 在低剂量的慢性单一和混合剂暴露后，在30天的少年阶段阶段鱼类中的PCB-11-硫酸盐毒性中的毒性。 执行这些目标的支撑培训计划包括学习和/或表演方法，例如Erod 测量Cyphnce，荧光显微镜以评估肝脏发育，组织病理学，油 - 红色-O染色以评估脂质沉积和荧光激活的细胞分选（FACS）脂肪酸 肝细胞的组成分析。 早期生命阶段（FELS）毒性测试（OECD TG 210）以获得调节剂实用经验。 采取措施并增加为政府胜任工作的职业目标的准备 污染物暴露的公共卫生影响。 PCB-11及其代谢物处于几个发展阶段，将是公共卫生监管机构。 赞助商Alicia Timme-Laragy博士拥有NIH资助的记录，是NIH F32的最新导师 博士后研究员（现在是圣地亚哥州立大学的教职员工），有许多设备 荧光显微镜，分子生物学和生物化学工作，能够提供指导 并需要完成该项目的指导。 研究所并拥有几种资源，使应用生命科学研究所（IALS）无关。 呢