Caruso J A, Stemmer P M
1 Proteomics Core Facility, Institute of Environmental Health Sciences, Wayne State University, Detroit, MI, USA.
2 Institute of Environmental Health Sciences, Wayne State University, Detroit, MI, USA.
Hum Exp Toxicol. 2018 Nov;37(11):1215-1232. doi: 10.1177/0960327118765326. Epub 2018 Mar 26.
Petroleum coke (PC) is a coal-like product that is produced during the refinement of crude oil and bituminous sand. Fugitive dust from open storage of PC in urban areas is a potential human health concern. Animal inhalation studies suggest that PC leads to an adverse pulmonary histopathology, including areas of fibrosis and chronic inflammation; however, little is known about its impact on human health. In order to identify biomarkers and cellular pathways that are associated with exposure, we performed two-dimensional liquid chromatography-mass spectrometric analyses on secreted proteins from two human lung culture models. A total of 2795 proteins were identified and relatively quantified from an immortalized cell line and 2406 proteins from primary cultures that were either mock treated or exposed to particulate matter with a diameter of 2.5-10 μm PC or filtered urban air particulates for 16 h. Pathway analysis on secretomes from primary lung cultures indicated that PC exposure suppressed the secretion of proteins involved in the organization of the extracellular matrix and epithelial differentiation. Because these cellular processes could facilitate fibrosis, we performed chronic 12-day exposure studies on three-dimensional human lung cultures consisting of epithelia and stromal fibroblasts. Relative to mock-treated cells, matrix metallopeptidase 9 levels in the conditioned media were lower by 4 days postexposure and remained suppressed for the duration of the experiment. Immunocytochemical staining of collagen III, a marker associated with fibrosis, showed increased accumulation in the epithelial layer and at the air-liquid interface.
石油焦(PC)是一种类似煤的产品,在原油和沥青砂的精炼过程中产生。城市地区露天储存石油焦产生的扬尘是一个潜在的人类健康问题。动物吸入研究表明,石油焦会导致肺部组织病理学不良变化,包括纤维化和慢性炎症区域;然而,人们对其对人类健康的影响知之甚少。为了确定与暴露相关的生物标志物和细胞途径,我们对两种人肺培养模型分泌的蛋白质进行了二维液相色谱-质谱分析。从永生化细胞系中总共鉴定并相对定量了2795种蛋白质,从原代培养物中鉴定并相对定量了2406种蛋白质,这些原代培养物要么进行了模拟处理,要么暴露于直径为2.5 - 10μm的石油焦颗粒物或过滤后的城市空气颗粒物中16小时。对原代肺培养物分泌组的通路分析表明,石油焦暴露抑制了参与细胞外基质组织和上皮分化的蛋白质的分泌。由于这些细胞过程可能促进纤维化,我们对由上皮细胞和基质成纤维细胞组成的三维人肺培养物进行了为期12天的慢性暴露研究。与模拟处理的细胞相比,暴露后4天条件培养基中基质金属肽酶9的水平降低,并且在实验期间一直受到抑制。与纤维化相关的标志物Ⅲ型胶原蛋白的免疫细胞化学染色显示,在上皮层和气液界面处的积累增加。