Mechanism of Triclosan Disruption of Mast Cell Function

三氯生破坏肥大细胞功能的机制

• 批准号: 8772031
• 负责人: Julie Ann Gosse
• 金额: $ 42.11万
• 依托单位: UNIVERSITY OF MAINE ORONO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8772031
• 关键词: Actins; Affect; Anti-Bacterial Agents; Antigens; Anxiety; Asthma; Autistic Disorder; Autoimmunity; Binding; Biochemical; Biological; Calcium Signaling; Cell Degranulation; Cell membrane; Cell physiology; Cells; Central Nervous System Diseases; Chemicals; Clinical Data; Communicable Diseases; Confocal Microscopy; Cytoplasmic Granules; Cytoskeleton; DNA Sequence Rearrangement; Data; Dextrans; Disease; Dose; Eczema; Elements; Environmental Exposure; Enzyme-Linked Immunosorbent Assay; Exposure to; F-Actin; Fluorescence; Fluorescence Microscopy; Hand; Health; Histamine Release; Hospitals; Household; Human; Hypersensitivity; IgE; IgE Receptors; Image; Inflammatory; Inflammatory Bowel Diseases; Intracellular Transport; Investigation; Ionophores; Knowledge; Laboratories; Life; Location; Malignant Neoplasms; Measures; Membrane; Methods; Microscopy; Microtubules; Molecular; Monitor; Multiple Sclerosis; Myosin ATPase; National Health and Nutrition Examination Survey; Neurons; Pharmacology and Toxicology; Phospholipase D; Physiological Processes; Population; Process; Protein Binding; Protein Isoforms; Protein Kinase C; Proteins; Public Health; Rattus; Reactive Oxygen Species; Reporter; Research; Resolution; Sampling; Self Care; Serotonin; Signal Pathway; Signal Transduction; Signaling Protein; Soaps; T-Lymphocyte; Triclosan; Tubulin; Urine; Work; antimicrobial; cell type; citrate carrier; dextran; effective therapy; insight; intercellular communication; mast cell; photoactivation; polymerization; public health relevance; receptor; research study; tool; toxicant

DESCRIPTION (provided by applicant): Triclosan (TCS) is an antimicrobial that is used widely in hospitals, consumer goods, and personal care products such as hand soaps at concentrations ~10 mM leading to wide exposure at high concentrations to consumers. Mast cells are critical players in numerous diseases, including allergy, asthma, autoimmunity, infectious disease, cancer, inflammatory bowel disease, and even many central nervous system disorders such as autism, anxiety, and multiple sclerosis. Due to the centrality of mast cells in myriad physiological processes and diseases and the ubiquitous exposure of the U.S. population to this chemical, there is an urgent need for information on the mammalian toxicology and pharmacology of TCS. The Gosse lab has shown that TCS inhibits both degranulation (in rat RBL-2H3 cells and human HMC-1 cells) and actin cytoskeletal rearrangement. Both of these processes are stimulated by both antigen via the Immunoglobulin E (IgE) receptor Fc?RI, and by Ca2+ ionophore. This project will investigate the cellular and molecular mechanisms by which TCS suppresses mast cell degranulation and F-actin ruffling via Protein Kinase C (PKC) and the actin cytoskeleton and also through Phospholipase D (PLD) and microtubule transport. The hypothesis is that TCS affects the activity and cellular localization of PKC and actin cytoskeleton. PKC activity will be measured through an ELISA. Initial confocal microscopy experiments will define TCS effects on general sub-cellular localization and dynamics of PKC, actin, and related proteins in live mast cells. Next, super-resolution imaging with fluorescence photoactivation localization microscopy (FPALM) will be used to investigate PKC:actin and other protein binding interactions not visible in traditional microscopy. Another hypothesis being studied is that TCS affects the activity and cellular localization of PLD and microtubule transport Using both confocal and FPALM, TCS effects on tubulin dynamics and PLD localization will be investigated. A new method that utilizes fluorescent dextran as a reporter will be used to image TCS effects on the final step of degranulation to directly quantify the locations and timeframes of degranulation. This research will fill in missing knowledge on the effects of TCS on mammalian signaling, and will allow prediction of TCS effects in disparate cell types that share common signal transduction elements. The results of this study will also fulfill an urgent need by providig insights into the impact of TCS on human health.

描述（由申请人提供）：三氯生 (TCS) 是一种抗菌剂，广泛用于医院、消费品和个人护理产品（例如洗手液），浓度约为 10 mM，导致消费者广泛接触高浓度。肥大细胞在许多疾病中发挥着重要作用，包括过敏、哮喘、自身免疫、传染病、癌症、炎症性肠病，甚至许多中枢神经系统疾病，如自闭症、焦虑症和多发性硬化症。由于肥大细胞在无数生理过程和疾病中的中心地位以及美国人普遍接触这种化学物质，因此迫切需要有关 TCS 的哺乳动物毒理学和药理学的信息。 Gosse 实验室表明，TCS 可以抑制脱颗粒（在大鼠 RBL-2H3 细胞和人 HMC-1 细胞中）和肌动蛋白细胞骨架重排。这两个过程均受到抗原通过免疫球蛋白 E (IgE) 受体 Fc?RI 和 Ca2+ 离子载体的刺激。该项目将研究 TCS 通过蛋白激酶 C (PKC) 和肌动蛋白细胞骨架以及磷脂酶 D (PLD) 和微管运输抑制肥大细胞脱颗粒和 F-肌动蛋白皱褶的细胞和分子机制。假设 TCS 影响 PKC 和肌动蛋白细胞骨架的活性和细胞定位。 PKC 活性将通过 ELISA 进行测量。最初的共聚焦显微镜实验将确定 TCS 对活体肥大细胞中 PKC、肌动蛋白和相关蛋白的一般亚细胞定位和动态的影响。接下来，采用荧光光激活定位显微镜 (FPALM) 的超分辨率成像将用于研究 PKC：肌动蛋白和其他传统显微镜中不可见的蛋白质结合相互作用。正在研究的另一个假设是 TCS 影响 PLD 和微管运输的活性和细胞定位。使用共聚焦和 FPALM，将研究 TCS 对微管蛋白动力学和 PLD 定位的影响。一种利用荧光右旋糖酐作为报告基因的新方法将用于成像 TCS 对脱颗粒最后一步的影响，以直接量化脱颗粒的位置和时间范围。 脱颗粒。这项研究将填补关于 TCS 对哺乳动物信号传导影响的知识缺失，并将允许预测 TCS 对共享共同信号转导元件的不同细胞类型的影响。这项研究的结果也将通过提供 TCS 对人类健康影响的见解来满足迫切的需求。

项目成果

Triclosan exposure, transformation, and human health effects.

三氯生暴露、转化和人类健康影响。

• DOI: 10.1080/10937404.2017.1399306
• 发表时间: 2017
• 期刊: Journal of toxicology and environmental health. Part B, Critical reviews
• 作者: Weatherly LM;Gosse JA
• 通讯作者: Gosse JA

Dances with Membranes: Breakthroughs from Super-resolution Imaging.

• DOI: 10.1016/bs.ctm.2015.03.008
• 发表时间: 2015
• 期刊: Current topics in membranes
• 作者: Curthoys NM;Parent M;Mlodzianoski M;Nelson AJ;Lilieholm J;Butler MB;Valles M;Hess ST
• 通讯作者: Hess ST

Triclosan is a mitochondrial uncoupler in live zebrafish.

• DOI: 10.1002/jat.3311
• 发表时间: 2016-12
• 期刊: JOURNAL OF APPLIED TOXICOLOGY
• 影响因子: 3.3
• 作者: Shim, Juyoung;Weatherly, Lisa M.;Luc, Richard H.;Dorman, Maxwell T.;Neilson, Andy;Ng, Ryan;Kim, Carol H.;Millard, Paul J.;Gosse, Julie A.
• 通讯作者: Gosse, Julie A.