Division of Infection, Immunity and Respiratory Medicine, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom.
NIHR-Manchester Biomedical Research Centre, Manchester University NHS Foundation Trust, Manchester, United Kingdom.
J Breath Res. 2023 Sep 7;17(4). doi: 10.1088/1752-7163/acf391.
Volatile organic compounds (VOCs) have shown promise as potential biomarkers in idiopathic pulmonary fibrosis. Measuring VOCs in the headspace ofmodels of lung fibrosis may offer a method of determining the origin of those detected in exhaled breath. The aim of this study was to determine the VOCs associated with two lung cell lines (A549 and MRC-5 cells) and changes associated with stimulation of cells with the pro-fibrotic cytokine, transforming growth factor (TGF)-1. A dynamic headspace sampling method was used to sample the headspace of A549 cells and MRC-5 cells. These were compared to media control samples and to each other to identify VOCs which discriminated between cell lines. Cells were then stimulated with the TGF-1 and samples were compared between stimulated and unstimulated cells. Samples were analysed using thermal desorption-gas chromatography-mass spectrometry and supervised analysis was performed using sparse partial least squares-discriminant analysis (sPLS-DA). Supervised analysis revealed differential VOC profiles unique to each of the cell lines and from the media control samples. Significant changes in VOC profiles were induced by stimulation of cell lines with TGF-1. In particular, several terpenoids (isopinocarveol, sativene and 3-carene) were increased in stimulated cells compared to unstimulated cells. VOC profiles differ between lung cell lines and alter in response to pro-fibrotic stimulation. Increased abundance of terpenoids in the headspace of stimulated cells may reflect TGF-1 cell signalling activity and metabolic reprogramming. This may offer a potential biomarker target in exhaled breath in IPF.
挥发性有机化合物（VOCs）已被证明是特发性肺纤维化潜在的生物标志物。测量肺纤维化模型中头空间中的 VOCs 可能提供一种确定在呼气中检测到的 VOCs 来源的方法。本研究的目的是确定与两种肺细胞系（A549 和 MRC-5 细胞）相关的 VOCs 以及与促纤维化细胞因子转化生长因子（TGF）-1 刺激细胞相关的变化。使用动态顶空采样方法来采样 A549 细胞和 MRC-5 细胞的头空间。将这些与培养基对照样品进行比较，并相互比较，以确定区分细胞系的 VOCs。然后用 TGF-1 刺激细胞，并在刺激和未刺激细胞之间比较样品。使用热解吸-气相色谱-质谱法分析样品，并使用稀疏偏最小二乘判别分析（sPLS-DA）进行监督分析。监督分析显示出每个细胞系特有的独特 VOC 图谱以及与培养基对照样品的差异。用 TGF-1 刺激细胞系会引起 VOC 图谱的显著变化。特别是，几种萜烯（异松油醇、菖蒲烯和 3-蒈烯）在刺激细胞中比未刺激细胞中增加。肺细胞系之间的 VOC 图谱不同，并对促纤维化刺激作出反应。刺激细胞头空间中萜烯的丰度增加可能反映了 TGF-1 细胞信号转导活性和代谢重编程。这可能为特发性肺纤维化的呼气中提供潜在的生物标志物靶标。
