Nephrotoxic mechanisms of f low-rank coal (lignite) leachates in drinking water

饮用水中流动煤（褐煤）渗滤液的肾毒性机制

• 批准号: 10082452
• 负责人: Michael A. Ihnat
• 金额: $ 7.25万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-08 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10082452
• 关键词: Address; Animals; Area; Arkansas; Balkan Nephropathy; Balkans; Bladder; Blood; Bosnia-Herzegovina; Brain; Bulgaria; Cell Death; Chemicals; Coal; Complete Blood Count; County; Creatinine; Croatia; Degenerative Disorder; Deposition; Disease; Dose; End stage renal failure; Environmental Risk Factor; Etiology; Exposure to; Female; Geography; Geology; Goals; Health; Heart; Hexanes; Hot Springs; Incidence; Inductively Coupled Plasma Mass Spectrometry; Inflammation; Kidney; Kidney Diseases; Link; Liver; Louisiana; Lung; Malignant Neoplasms; Mass Fragmentography; Mediating; Metals; Methanol; Microscopic; Nature; Oklahoma; Organ; Osmolalities; Pathologist; Phase; Process; Publishing; Rattus; Recording of previous events; Renal function; Romania; Rural; Sampling; Serbia; Serum; Skin; Solid; Spleen; Sprague-Dawley Rats; Testing; Texas; Toxic effect; Toxicokinetics; Universities; Urea Nitrogen; Urine; Urothelium; Water consumption; Weight; Work; aqueous; cytotoxic; cytotoxicity; drinking water; graduate student; gulf coast; in vivo; kidney cell; male; nephrotoxicity; renal damage; undergraduate student

Abstract Balkan endemic nephropathy (BEN) is a degenerative disease that is often associated with urothelial cancer and is geographically confined to rural parts of Serbia, Bulgaria, Romania, Croatia, and Bosnia. The etiology of BEN is debated, and several environmental factors have been considered as contributors to the disease. The Pliocene lignite hypothesis links BEN and exposure to organic material in drinking water that is leached from a low-rank coal known as lignite. This hypothesis addresses the limited geographic nature of BEN, as most of the villages plagued with the disease are also closely associated with lignite deposits. We and others have shown an association of lignite deposits in the southeastern US with renal cell toxicity in culture and with an increased incidence of end-stage renal disease (ESRD).The overall goal of the studies in this work are to test the specific hypothesis that environmental levels of U.S. Gulf Coast region lignite aqueous extracts in drinking water result in BEN-like nephrotoxicity in vivo. Aim 1 will test the prediction that drinking water exposure to lignite coal extracts from the southeastern US results in a dose-dependent nephrotoxicity. In this study, male and female Sprague-Dawley (S-D) rats will be exposed to environmental levels of coal extracts in their drinking water for six months. Samples will be either lignite extracts found to be toxic to renal cells (Dolet Hills, LA and Monticello, TX); or lignite extracts found to be less toxic to kidney cells (San Miguel, TX; Hot Springs County, AR). Water intake, animal weights and general health will be assessed weekly. Blood and 24 hr urine will be collected at 3 and 6 months from a subset of animals and assessed for renal function markers (urine osmolality, NAG levels; serum urea nitrogen and creatinine). Kidneys will be assessed for toxicity, cell death and inflammation grossly and microscopically by a veterinary pathologist as will other potential target organs (bladder, lungs, heart, liver, spleen, brain, skin, complete blood count). Aim 2 will test the prediction that lignite degradation products, specifically polar organic compounds, are cytotoxic to renal cells. In this aim, toxic extracts (Dolet Hills, LA; Monticello, TX) will be separated by solid-phase extraction followed by a two-step elution process to generate a non-polar component (compounds soluble in hexane) and polar component (compounds soluble in methanol). These fractions applied at various concentrations as in our preliminary work to HK-2 and RPTEC/TERT1 renal cells and assessed for cytotoxicity. The impact of this proposal is to begin to tie these two findings together, namely to test whether lignite coal extracts in the drinking water of rats is nephrotoxic and to begin to understand the lignite components/mechanisms mediating this nephrotoxicity.

摘要 巴尔干半岛的地方性肾病（BEN）是一种经常与尿路上皮癌相关的退行性疾病， 在地理上仅限于塞尔维亚、保加利亚、罗马尼亚、克罗地亚和波斯尼亚的农村地区。BEN的病因 有争议的是，几个环境因素被认为是疾病的贡献者。上新世 褐煤假说将BEN和暴露于饮用水中的有机物质联系起来，饮用水是从低等级的 煤称为褐煤。这一假设解决了本恩有限的地理性质，因为大多数那些村庄 与疾病的困扰也密切相关的褐煤矿床。我们和其他人已经展示了一个 美国东南部的褐煤沉积物与培养物中的肾细胞毒性以及 终末期肾病（ESRD）的发病率。这项工作中研究的总体目标是测试特定的 假设美国墨西哥湾沿岸地区饮用水中褐煤水提取物的环境水平导致 体内BEN样肾毒性。目标1将检验饮用水暴露于褐煤的预测 来自美国东南部的提取物导致剂量依赖性肾毒性。在这项研究中，男性和女性 Sprague-Dawley（S-D）大鼠将暴露于其饮用水中的煤提取物的环境水平， 个月样本将是发现对肾细胞有毒的褐煤浸提液（Dolet Hills，LA和蒙蒂塞洛， TX）;或发现对肾细胞毒性较小的褐煤提取物（San Miguel，TX; Hot Springs County，AR）。水 每周评估摄入量、动物体重和一般健康状况。血液和24小时尿液将在3时采集 和6个月，并评估肾功能标志物（尿渗透压，NAG水平; 血清尿素氮和肌酸酐）。将对肾脏的毒性、细胞死亡和炎症进行大体评估 以及其它潜在的靶器官（膀胱，肺，心脏，肝， 脾、脑、皮肤、全血细胞计数）。目标2将测试褐煤降解产物的预测， 特别是极性有机化合物对肾细胞具有细胞毒性。在这个目标中，有毒提取物（Dolet Hills，LA; 蒙蒂塞洛，德克萨斯州）将通过固相萃取分离，然后进行两步洗脱过程， 非极性组分（可溶于己烷的化合物）和极性组分（可溶于甲醇的化合物）。 如我们对HK-2和RPTEC/TERT 1肾细胞的初步工作中，这些组分以不同浓度应用 细胞并评估细胞毒性。这一提议的影响是开始将这两个发现联系在一起， 即测试大鼠饮用水中的褐煤提取物是否具有肾毒性，并开始了解 介导这种肾毒性的褐煤组分/机制。