Van Den Broucke Sofie, Vanoirbeek Jeroen A J, Derua Rita, Hoet Peter H M, Ghosh Manosij
Centre for Environment and Health, Department of Public Health and Primary Care, KU Leuven, 3000 Leuven, Belgium.
Laboratory of Respiratory Diseases and Thoracic Surgery (BREATHE), KU Leuven, 3000 Leuven, Belgium.
Nanomaterials (Basel). 2021 May 13;11(5):1283. doi: 10.3390/nano11051283.
Via inhalation we are continuously exposed to environmental and occupational irritants which can induce adverse health effects, such as irritant-induced asthma (IIA). The airway epithelium forms the first barrier encountered by these agents. We investigated the effect of environmental and occupational irritants on the airway epithelial barrier in vitro. The airway epithelial barrier was mimicked using a coculture model, consisting of bronchial epithelial cells (16HBE) and monocytes (THP-1) seeded on the apical side of a permeable support, and human lung microvascular endothelial cells (HLMVEC) grown on the basal side. Upon exposure to graphene (G) and graphene oxide (GO) in a suspension with fetal calf serum (FCS), ammonium persulfate (AP), sodium persulfate (SP) and hypochlorite (ClO), the transepithelial electrical resistance (TEER) and flux of fluorescent labelled dextran (FD4-flux), was determined. Exposure to graphene nanoparticles (GNPs) induced an immediate negative effect on the epithelial barrier, whereas ClO only had a negative impact after 24 h of exposure. AP and SP did not affect the barrier properties. The tight junctions (TJ) network showed less connected zonula occludens 1 (ZO-1) and occludin staining in GNP-exposed cocultures. Functional analysis of the phosphoproteomic data indicated that proteins in the adherens junction (AJ) and TJ pathways showed an altered phosphorylation due to GNP exposure. To conclude, the negative effect of GNPs on the epithelial barrier can be explained by the slightly altered the TJ organization which could be caused by alterations in the phosphorylation level of proteins in the AJ and TJ pathway.
通过吸入，我们持续接触环境和职业性刺激物，这些刺激物可引发不良健康影响，如刺激性哮喘（IIA）。气道上皮是这些物质遇到的第一道屏障。我们在体外研究了环境和职业性刺激物对气道上皮屏障的影响。使用共培养模型模拟气道上皮屏障，该模型由接种在可渗透支持物顶侧的支气管上皮细胞（16HBE）和单核细胞（THP-1）以及生长在基底侧的人肺微血管内皮细胞（HLMVEC）组成。在含有胎牛血清（FCS）的悬浮液中暴露于石墨烯（G）、氧化石墨烯（GO）、过硫酸铵（AP）、过硫酸钠（SP）和次氯酸盐（ClO）后，测定跨上皮电阻（TEER）和荧光标记葡聚糖通量（FD4-通量）。暴露于石墨烯纳米颗粒（GNP）对上皮屏障立即产生负面影响，而ClO仅在暴露24小时后有负面影响。AP和SP不影响屏障特性。紧密连接（TJ）网络在暴露于GNP的共培养物中显示闭合蛋白1（ZO-1）和闭合蛋白染色的连接减少。磷酸化蛋白质组学数据的功能分析表明，由于GNP暴露，黏着连接（AJ）和TJ途径中的蛋白质磷酸化发生改变。总之，GNP对上皮屏障的负面影响可通过TJ组织的轻微改变来解释，这可能是由AJ和TJ途径中蛋白质磷酸化水平的改变引起的。
