College of Pharmacy, Binzhou Medical University, Yantai, 264003, Shandong, China.
Zhangjiang Lab, Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai, 201210, China.
Anal Bioanal Chem. 2019 Nov;411(27):7157-7164. doi: 10.1007/s00216-019-02083-8. Epub 2019 Sep 6.
Endothelial damage is a major manifestation in many forms of heart and lung injuries induced by lipopolysaccharide (LPS), but the biochemical responses and activation mechanisms of endothelial cells have not been fully explicit. In this study, the biochemical changes to endothelial cells exposed to LPS were investigated by synchrotron FTIR microspectroscopy at a single-cell level. We found that the whole infrared spectrum of endothelial cells shifted after LPS treatment, indicating chemical component changes within cells. Principal component analysis (PCA) and t tests on subspectra (fatty acid region, protein region, and nucleic acid-sugar region, respectively) further showed that sugar components as well as fatty acids changed dramatically while proteins had no significant variation following LPS exposure. These results suggested that the glycocalyx layer structure on endothelial cell membrane may be mainly influenced by LPS and also proved that synchrotron FTIR microspectroscopy was a useful technique to evaluate the biochemical changes of endothelial damage at the single-cell level. Graphical abstract.
内皮细胞损伤是脂多糖（LPS）诱导的多种心肺损伤的主要表现形式，但内皮细胞的生化反应和激活机制尚未完全阐明。本研究采用同步辐射傅里叶变换红外（FTIR）微光谱技术在单细胞水平上研究了 LPS 作用下内皮细胞的生化变化。我们发现 LPS 处理后内皮细胞的整个红外光谱发生了位移，表明细胞内化学组成发生了变化。主成分分析（PCA）和子谱（脂肪酸区、蛋白质区和核酸-糖区）的 t 检验进一步表明，糖成分以及脂肪酸在 LPS 暴露后发生了显著变化，而蛋白质没有明显变化。这些结果表明，LPS 可能主要影响内皮细胞膜糖萼层的结构，并证明同步辐射 FTIR 微光谱技术是评估内皮损伤生化变化的有用技术。
