石家庄市空气pm2.5对人肺泡上皮细胞a549的 氧化损伤研究-oxidative damage of air pm 2.5 to human alveolar epithelial cell a549 in shijiazhuang city.docxVIP

目录中文摘要···········································································1英文摘要···········································································3英文缩写···········································································6研究论文石家庄市空气PM2.5对人肺泡上皮细胞A549的氧化损伤研究前言···········································································7材料与方法································································8结果········································································16附图········································································19附表········································································23讨论········································································32结论········································································36参考文献··································································37综述雾霾对人群健康影响概述··········································41致谢···········································································51个人简历········································································52石家庄市空气PM2.5对人肺泡上皮细胞A549的氧化损伤研究摘要目的：研究大气细颗粒污染物PM2.5导致人肺泡上皮细胞损伤的主要作用，以及染毒浓度和染毒时间对肺上皮细胞毒性损伤的交互效应，为大气雾霾治理和人群呼吸系统疾病预防提供可靠的科学依据。方法：使用EP-100智能PM2.5采样器收集PM2.5颗粒物至玻璃纤维滤膜，将采集后的滤膜用超纯水超声洗脱，然后收集洗脱液，通过真空冷冻干燥机干燥，获取颗粒物后，使用PBS和F12培养基按一定比例配成浓度分别为0、25、50、100、200和400μg/ml的染毒液，使A549细胞暴露于各浓度染毒液，分别培养6h、24h和48h。染毒结束后收集上清液以及超声裂解的细胞裂解液。采用微量酶标法检测上清液中乳酸脱氢酶（LDH）漏出量，可见光法测定细胞内CAT活性，TBA法检测细胞膜丙二醛（MDA）生成量，羟胺法测细胞内超氧化物歧化酶（SOD）活性。应用Excel2013汇总实验数据和绘制图表，SPSS21.0分析软件处理和分析数据。Dunnett-t检验法用于分析染毒24h和48h组的毒性指标变化分别与6h组的差异，以及其余5个染毒浓度组的毒性指标变化分别与0μg/ml组的差异。单因素方差分析法用于探讨相同染毒时间内不同染毒浓度组之间以及相同染毒浓度下不同染毒时间组之间细胞毒性指标变化的差异，析因方差分析用于探讨染毒浓度和染毒时间对细胞产生毒性损伤的交互效应。结果：1细胞毒性指标变化趋势：不同染毒浓度对A549细胞的毒性损伤表现为LDH漏出量随PM2.5染毒浓度增加而增加，由低到高依次为：79.45、103.38、115.24、127.69、142.53和175.81U/gprot。不同染毒时间对A549细胞的毒性损伤为LDH漏出量随染毒时间延长而增加：染毒48h（167.28U/gprot）24h（109.92U/gprot）6h（94.86U/gprot）。MDA生成量随染毒浓度和染毒时间的变化趋势与LDH的一致。而CAT和SOD活性则是随着染毒浓度的增加或染毒时间的延长而降低。2不同染毒浓度的毒性作用：Dunnett-t检验结果显示，同一染毒时间组