Evaluating the biochemical changes of LPS-stimulated endothelial cells by synchrotron FTIR microspectroscopy at a single-cell level.

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.