Evaluating the biochemical changes of LPS-stimulated endothelial cells by synchrotron FTIR microspectroscopy at a single-cell level
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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.
KeywordsEndothelial cells Lipopolysaccharide Synchrotron FTIR microspectroscopy Single-cell analysis Glycocalyx
We thank the staff of BL01B beamline at the National Center for Protein Sciences Shanghai and Shanghai Synchrotron Radiation Facility for assistance during data collection.
This work was supported by the National Natural Science Foundation of China (No U1732130), Key Research Program of Frontier Sciences of the Chinese Academy of Sciences (QYZDJ-SSW-SLH019), and the Scientific Research Foundation of Binzhou Medical University (BY2017KYQD02).
Compliance with ethical standards
Conflict of interest
The authors declare that there are no conflicts of interest.
- 3.Stasi A, Intini A, Divella C, Franzin R, Montemurno E, Grandaliano G, et al. Emerging role of lipopolysaccharide binding protein in sepsis-induced acute kidney injury. Nephrol Dial Transplant. 2017;32(1):24–31.Google Scholar
- 7.Marechal X, Favory R, Joulin O, Montaigne D, Hassoun S, Decoster B, et al. Endothelial glycocalyx damage during endotoxemia coincides with microcirculatory dysfunction and vascular oxidative stress. Shock. 2008;29(5):572–6.Google Scholar
- 8.Xiao JG, Song Q, Li TS, Sun RJ. Bacterial endotoxin-induced endothelial cell injury and calcium overload associated with Toll-like receptor and calcium signal. Chin Crit Care Med. 2017;29(2):150–5.Google Scholar
- 26.Kamnev AA, Tugarova AV, Dyatlova YA, Tarantilis PA, Grigoryeva OP, Fainleib AM, et al. Methodological effects in Fourier transform infrared (FTIR) spectroscopy: implications for structural analyses of biomacromolecular samples. Spectrochim Acta A Mol Biomol Spectrosc. 2018;193:558–64.CrossRefGoogle Scholar
- 28.Zimkus A, Misiūnas A, Chaustova L. Li+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FTIR spectroscopy. Cent Euro J Bio. 2013;8(8):724–9.Google Scholar