IRON, CALCIUM AND OXIDATIVE STRESS IN LUNG INJURY

肺损伤中的铁、钙和氧化应激

• 批准号: 6445972
• 负责人: XI HUANG
• 金额: $ 32.43万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2003-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6445972
• 关键词: X ray crystallography; air sampling /monitoring; animal tissue; arsenic; cadmium; calcium; cell line; coal; cobalt; copper; dust; environmental contamination; ferritin; fibronectins; guinea pigs; human tissue; interleukin 1; iron; lung injury; macrophage; metallothionein; minings; nickel; occupational health /safety; oxidative stress; peroxidation; statistics /biometry; tissue /cell culture; tumor necrosis factor alpha; zinc

Striking differences in the prevalence of coal workers' pneumoconiosis and chronic obstructive bronchopneumopathy exist between different coal mine regions despite comparable levels of dust exposure. The goal of this proposal is to test the hypothesis that bio-available Fe2+ is the active component in coal dust-induced cell injury through oxidative stress pathway, and that calcite (CaCO) may play a protective role in such injury by inhibiting solubilization of Fe2+, making Fe2+ less bio- available. Primary cultures of guinea pig alveolar macrophages (AMs) and human lung type II cells (A549) will be used for coal treatments. Total of thirty coal samples from three coal mine regions with a CWP prevalence of 4 percent (Utah), 10 percent (West Virginia), and 26 percent (Pennsylvania), respectively, will be tested. The bio-availability of Fe2+ and Ca2+, as well as other trace metals (i.e. As, Cd, Ni, Zn, Co, Cu) and quartz released from coal dusts in these cells will be determined. Ferritin and metallothionein will be measured as indices of the bio-availability of Fe2+ and trace metals in these cells. Cellular Fe2+ and Ca2+ levels will also be determined using colorimetric methods. Quartz content in the cells will be quantitated using X-ray diffraction after low temperature ashing of cell matrix. Lipid peroxidation of cell membranes, H2O2 formation, and AP-1 expression will be measured before and after treatment with coals. The levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), and fibronectin will also be determined as a part of this study. Using TNF-alpha, IL-1, and other biochemical parameters as independent variables, the independent effect of bio-available Fe2+, trace metals, and quartz will be identified by multiple regression analyses. A pre-determined percentage of calcite will be added to the Pennsylvania and West Virginia coals before cell treatment. AMs and A549 cells will also be pretreated with Ca2+ before exposure to the untreated coals. The above mentioned end-points will be measured and effects of treatments assessed. We expect that, by confirming the role of Fe2+ in coal-induced injury, we will be able to predict which coal is likely to be more toxic, even before mining. By determining the role of calcite, it is possible to propose alternative methods for lung disease prevention.

尽管接尘水平相当，但不同矿区煤工尘肺和慢性阻塞性支气管肺疾病的患病率存在显著差异。 本提案的目的是验证以下假设：生物可利用的Fe 2+是通过氧化应激途径引起的细胞损伤中的活性组分，并且方解石（CaCO）可能通过抑制Fe 2+的增溶，使Fe 2+生物可利用性降低，在这种损伤中发挥保护作用。 将使用豚鼠肺泡巨噬细胞（AM）和人肺II型细胞（A549）的原代培养物进行煤炭处理。 将对来自三个煤矿区的总共30个煤样品进行测试，其中CWP患病率分别为4%（犹他州）、10%（西弗吉尼亚州）和26%（宾夕法尼亚州）。 将确定这些细胞中煤尘释放的Fe 2+和Ca 2+以及其他微量金属（即As、Cd、Ni、Zn、Co、Cu）和石英的生物可利用性。铁蛋白和金属硫蛋白将作为这些细胞中Fe 2+和痕量金属的生物利用度的指标进行测量。还将使用比色法测定细胞Fe 2+和Ca 2+水平。 细胞基质低温灰化后，使用X射线衍射定量细胞中的石英含量。 在用煤处理之前和之后测量细胞膜的脂质过氧化、H2 O2形成和AP-1表达。 作为本研究的一部分，还将测定肿瘤坏死因子-α（TNF-α）、白细胞介素-1（IL-1）和纤连蛋白的水平。使用TNF-α、IL-1和其他生化参数作为自变量，通过多元回归分析确定生物可利用的Fe 2+、微量金属和石英的独立影响。 在电池处理之前，将预定百分比的方解石添加到宾夕法尼亚和西弗吉尼亚的煤中。 AM和A549细胞在暴露于未处理的煤之前也将用Ca 2+预处理。 将测量上述终点并评估治疗效果。 我们希望，通过确认Fe 2+在煤引起的伤害中的作用，我们将能够预测哪种煤可能更具毒性，甚至在开采之前。 通过确定方解石的作用，有可能提出预防肺部疾病的替代方法。