PDF(Pubmed)
Abstract：
Air pollutants are associated with exacerbations of asthma, chronic bronchitis, and airway inflammation. Diesel exhaust particles (DEPs) can induce and worsen lung diseases. However, there are insufficient data to guide polymerase chain reaction (PCR) array proteomics studies regarding the impacts of DEPs on respiratory diseases. This study was performed to identify genes and proteins expressed in normal human bronchial epithelial (NHBE) cells. MicroRNAs (miRNAs) and proteins expressed in NHBE cells exposed to DEPs at 1 μg/cm2 for 8 h and 24 h were identified using PCR array analysis and 2D PAGE/LC-MS/MS, respectively. YWHAZ gene expression was estimated using PCR, immunoblotting, and immunohistochemical analyses. Genes discovered through an overlap analysis were validated in DEP-exposed mice. Proteomics approaches showed that exposing NHBE cells to DEPs led to changes in 32 protein spots. A transcriptomics PCR array analysis showed that 6 of 84 miRNAs were downregulated in the DEP exposure groups compared to controls. The mRNA and protein expression levels of YWHAZ, β-catenin, vimentin, and TGF-β were increased in DEP-treated NHBE cells and DEP-exposed mice. Lung fibrosis was increased in mice exposed to DEPs. Our combined PCR array-omics analysis demonstrated that DEPs can induce airway inflammation and lead to lung fibrosis through changes in the expression levels of YWHAZ, β-catenin, vimentin, and TGF-β. These findings suggest that dual approaches can help to identify biomarkers and therapeutic targets involved in pollutant-related respiratory diseases.
摘要：
空气污染物与哮喘的恶化有关,慢性支气管炎,和气道炎症。柴油机废气颗粒（DEP）可诱发和加重肺部疾病。然而,没有足够的数据指导有关DEP对呼吸系统疾病影响的聚合酶链反应(PCR)阵列蛋白质组学研究.进行这项研究以鉴定在正常人支气管上皮（NHBE）细胞中表达的基因和蛋白质。使用PCR阵列分析和2DPAGE/LC-MS/MS鉴定暴露于1μg/cm2DEP8小时和24小时的NHBE细胞中表达的微小RNA（miRNA）和蛋白质。分别。使用PCR估计YWHAZ基因表达，免疫印迹,和免疫组织化学分析。通过重叠分析发现的基因在暴露于DEP的小鼠中得到验证。蛋白质组学方法表明，将NHBE细胞暴露于DEP导致32个蛋白质点的变化。转录组学PCR阵列分析显示,与对照相比,在DEP暴露组中84个miRNA中的6个下调。YWHAZ的mRNA和蛋白表达水平,β-连环蛋白，波形蛋白,在DEP处理的NHBE细胞和暴露于DEP的小鼠中TGF-β增加。暴露于DEP的小鼠肺纤维化增加。我们的联合PCR阵列-组学分析表明，DEP可以通过改变YWHAZ的表达水平来诱导气道炎症并导致肺纤维化，β-连环蛋白，波形蛋白,和TGF-β。这些发现表明，双重方法可以帮助识别与污染物相关的呼吸系统疾病有关的生物标志物和治疗靶标。
