内蒙古白云鄂博稀土矿区大气可吸入颗粒物对A549细胞毒理研究

项目名称： 内蒙古白云鄂博稀土矿区大气可吸入颗粒物对A549细胞毒理研究

项目编号： No.81473017

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 孙涓

作者单位： 内蒙古医科大学

项目金额： 66万元

中文摘要： 本研究拟对内蒙古白云鄂博稀土矿区进行空气颗粒物（PMs）取样。超声法提取PMs并配置成不同浓度，运用ICP-MS和GC-MS分析其化学组成。以人肺腺癌A549细胞为模型，采用MTT法测定细胞活力。以稀土矿区上风向空气PMs为对照，观察下风向区PMs对A549细胞活力的影响；采用流式细胞仪观察矿区PMs对A549细胞的影响，检测PMs对A549细胞活性氧水平的影响；采用荧光显微镜观察PMs对A549细胞的凋亡形态学的影响，并对细胞内Caspase3/6/8/9以及NF-kβ(p65)表达量进行检测；釆用蛋白质组学方法比较不同区域PMs对A549细胞蛋白表达谱的差异，得出空气PMs中稀土元素潜在的毒性作用。筛选干预A549细胞的过程中差异表达的蛋白及它们参与的生物过程，分析差异蛋白与A549细胞周期、细胞凋亡之间的关系.为评价稀土矿区环境颗粒物对呼吸系统危害提供基础数据。

中文关键词： 可吸入颗粒物；蛋白质组学；A549细胞；稀土；细胞毒性

英文摘要： This Study intends to collecte particulate matters in the rare earths mine area of Bayan Obo, Inner Mongolia.The total PMs extracted by ultrasonic method and dissolved in different concentrations. Contents of chemical composition determined by ICP-MS and GC-MS. A549 cells model applied which simulate the process of in vivo exposure.Cytotoxicity of PMs measured by MTT assay，compared with PMs in upwind area and PMs in downwind area on the A549 cytotoxicity；to examine whether PMs could influence the cell cycle progression, FACS performed, the intracelluar reactive oxygen species (ROS) levels detected as well；the morphological changes of apoptosis of A549 cells observed by the fluorescence microscopy and the intracellular activity of Caspase3/6/8/9 and the content of NF-kappa β(p65) detected；the protein expression profile of A549 cells after intervention by PMs analyized by the proteomics technology, which would provide some information about the differentially expressed proteins,and understand potential toxic effects of rare earths elements in PMs. Differential express protein and biological processes for this proteins of A549 cell after treated by PMs, and analyze the relationship between differentially expressed proteins，cell cycle and apoptosis. On the basis of monitoring environmental PMs and pulmonary function for exposed participants in rare earths mine area.

英文关键词： particulate matters;proteome;A549 cell;rare earths;cytotoxicity