IRON, CALCIUM AND OXIDATIVE STRESS IN LUNG INJURY

肺损伤中的铁、钙和氧化应激

• 批准号: 6042278
• 负责人: XI HUANG
• 金额: $ 31.83万
• 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 2002-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6042278
• 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，但不同煤矿地区煤工尘肺和慢性阻塞性肺疾病的患病率存在显着差异。该方案的目的是验证生物有效Fe2+是煤尘通过氧化应激途径引起的细胞损伤中的活性成分的假说，以及方解石(CaCO)可能通过抑制Fe2+的增溶而减少Fe2+的生物可利用性而在这种损伤中起到保护作用。原代培养的豚鼠肺泡巨噬细胞(AM)和人肺II型细胞(A549)将用于煤炭治疗。来自三个煤粉病患病率分别为4%(犹他州)、10%(西弗吉尼亚州)和26%(宾夕法尼亚州)的煤矿地区的30个煤炭样本将进行测试。将测定煤尘中释放出的Fe2+、Ca2+以及其他痕量金属(As、Cd、Ni、Zn、Co、Cu)和石英在这些单元中的生物有效性。铁蛋白和金属硫蛋白将作为这些细胞中Fe2+和痕量金属的生物有效性的指标进行测量。细胞内的Fe2+和Ca2+水平也将用比色法测定。电池基质低温灰化后，用X射线衍射法测定电池中的石英含量。用煤处理前后将检测细胞膜的脂质过氧化、H_2O_2的形成和AP-1的表达。肿瘤坏死因子-α(TNF-α)、白介素1(IL-1)和纤维连接蛋白的水平也将作为本研究的一部分进行测定。以肿瘤坏死因子-α、白介素1等生化参数为自变量，通过多元回归分析确定生物有效铁、痕量金属和石英的独立效应。在单元处理之前，宾夕法尼亚州和西弗吉尼亚州的煤炭中将添加预定比例的方解石。AMS和A549细胞在接触未经处理的煤炭之前也将被钙离子预处理。将测量上述终点并评估治疗效果。我们预计，通过确认Fe2+在煤炭伤害中的作用，我们将能够预测哪些煤炭可能更具毒性，甚至在开采之前就能预测。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。