Effect of sub-chronic exposure to cigarette smoke, electronic cigarette and waterpipe on human lung epithelial barrier function
Effect of sub-chronic exposure to cigarette smoke, electronic cigarette and waterpipe on human lung epithelial barrier function
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DOI：
10.1186/s12890-020-01255-y
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发表时间：
2020-08-12
影响因子：
3.1
通讯作者：
Sidhaye, Venkataramana K.
中科院分区：
文献类型：
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作者：
Ghosh, Baishakhi;Reyes-Caballero, Hermes;Sidhaye, Venkataramana K.
背景考虑到最近与电子烟使用相关的肺损伤病例激增，我们设计了人支气管上皮细胞（HBEC）在气液界面中亚慢性暴露的体外模型，以确定亚慢性暴露于香烟烟雾（CS）、电子烟气溶胶（EC）和烟草水烟暴露（TW）导致的上皮细胞屏障的恶化。方法分析的产品包括市售电子烟油，浓度为0%或1.2% 浓度的尼古丁、烟草混合物（水烟）和参考级卷烟 (3R4F)。在一组实验中，使用 1 支香烟/天将 HBEC 暴露于 EC（0 和 1.2%）、CS 或对照空气中 10 天。在第二组实验中，将假复层原代上皮组织暴露于TW或对照空气暴露中，每隔一天进行1小时，直到进行3次暴露。最后一次暴露 16-18 小时后，我们使用异硫氰酸荧光素-葡聚糖通量测定 (FITC-Dextran)、跨电上皮电阻 (TEER) 测量、运动纤毛百分比评估、纤毛跳动频率 (CBF)、细胞运动以及通过反转录定量聚合酶链反应对 E-钙粘蛋白基因表达进行定量来研究上皮单层的屏障功能/结构完整性(RT-qPCR)。结果与空气对照相比，CS 使荧光（FITC-葡聚糖测定）增加了 5.6 倍，其中 CS 和 EC (1.2%) 分别将 TEER 降低至 49% 和 60%。与对照空气相比，CS 和 EC (1.2%) 暴露使 CBF 分别降低至 62% 和 59%，纤毛移动分别降低至 47% 和 52%。与空气对照相比，CS 和 EC (1.2%) 的细胞速度分别提高了 2.5 和 2.6 倍。通过 CS 暴露，E-钙粘蛋白的表达降低至对照空气水平的 39%，这表明对合理的分子机制的深入了解。总而言之，EC (0%) 和 TW 暴露导致上皮完整性的中度下降，而 EC (1.2%) 与 CS 暴露相比显着降低了气道上皮屏障功能。结论结果支持亚慢性暴露于 EC (1.2%) 的毒性作用，通过破坏支气管上皮细胞屏障完整性来证明这一点，而需要进一步研究来解决这一观察结果的分子机制以及长期暴露中 TW 和 EC (0%) 的毒性。
BackgroundTaking into consideration a recent surge of a lung injury condition associated with electronic cigarette use, we devised an in vitro model of sub-chronic exposure of human bronchial epithelial cells (HBECs) in air-liquid interface, to determine deterioration of epithelial cell barrier from sub-chronic exposure to cigarette smoke (CS), e-cigarette aerosol (EC), and tobacco waterpipe exposures (TW).MethodsProducts analyzed include commercially available e-liquid, with 0% or 1.2% concentration of nicotine, tobacco blend (shisha), and reference-grade cigarette (3R4F). In one set of experiments, HBECs were exposed to EC (0 and 1.2%), CS or control air for 10days using 1 cigarette/day. In the second set of experiments, exposure of pseudostratified primary epithelial tissue to TW or control air exposure was performed 1-h/day, every other day, until 3 exposures were performed. After 16-18h of last exposure, we investigated barrier function/structural integrity of the epithelial monolayer with fluorescein isothiocyanate-dextran flux assay (FITC-Dextran), measurements of trans-electrical epithelial resistance (TEER), assessment of the percentage of moving cilia, cilia beat frequency (CBF), cell motion, and quantification of E-cadherin gene expression by reverse-transcription quantitative polymerase chain reaction (RT-qPCR).ResultsWhen compared to air control, CS increased fluorescence (FITC-Dextran assay) by 5.6 times, whereby CS and EC (1.2%) reduced TEER to 49 and 60% respectively. CS and EC (1.2%) exposure reduced CBF to 62 and 59%, and cilia moving to 47 and 52%, respectively, when compared to control air. CS and EC (1.2%) increased cell velocity compared to air control by 2.5 and 2.6 times, respectively. The expression of E-cadherin reduced to 39% of control air levels by CS exposure shows an insight into a plausible molecular mechanism. Altogether, EC (0%) and TW exposures resulted in more moderate decreases in epithelial integrity, while EC (1.2%) substantially decreased airway epithelial barrier function comparable with CS exposure.ConclusionsThe results support a toxic effect of sub-chronic exposure to EC (1.2%) as evident by disruption of the bronchial epithelial cell barrier integrity, whereas further research is needed to address the molecular mechanism of this observation as well as TW and EC (0%) toxicity in chronic exposures.