Abstract
Endothelial HMEC-1 cells incubated with pro-inflammatory cytokine TNF-α for 6 and 24 hours were studied as a model of inflammation using Raman imaging. Striking changes in distribution, composition and concentration of cellular lipids were observed after exposure to TNF-α compared to the control. In particular, 3D Raman imaging revealed a significant increase in the amount of lipid entities formed under inflammation. Lipid bodies were randomly distributed in the cytoplasm and two types of droplets were assembled: more saturated one, in spectral characteristics resembling phosphatidylcholine and saturated cholesteryl esters, observed also in the control, and highly unsaturated one, containing also cholesterols, being a hallmark of inflamed cells. The statistical analysis showed that the number of lipid bodies was significantly dependent on the exposure time to TNF-α. Overall, observed formation of unsaturated lipid droplets can be directly correlated with the increase in production of prostacyclins - endogenous inflammation mediators.
Highlights
Endothelial HMEC-1 cells incubated with pro-inflammatory cytokine TNF-α for 6 and 24 hours were studied as a model of inflammation using Raman imaging
The growing evidence indicates that some cardiovascular events and lifestyle diseases i.e. atherosclerosis, diabetes and hypertension begin with endothelial dysfunction[1,2,3,4,5] and thereby, endothelial cell cultures are convenient models to study pathology development in the circulatory system
Only few reports have been previously published regarding human endothelial cells lines cultures studied by means of Raman microscopy[6,7,8,9,10,11]
Summary
Endothelial HMEC-1 cells incubated with pro-inflammatory cytokine TNF-α for 6 and 24 hours were studied as a model of inflammation using Raman imaging. The possibility to obtain 3D Raman images of cells enables to visualize the distribution and track of newly formed entities inside the cell body This approach can compete with confocal fluorescence imaging where the number of simultaneously analyzed components is limited. TNF-αmay activate a number of signaling pathways in endothelial cells and the main consequences of its action is an increase in the intercellular adhesion molecules secretion and leucocyte adhesion[34], alterations of nitric oxide expression correlated with elasticity[35], the increase of reactive oxygen species (ROS) production[33] and apoptosis[36]. Raman imaging enabled characterization of composition of lipid bodies formed upon inflammation, while 3D profiling showed their exact intercellular distribution. Lipid droplets may become a universal Raman marker of inflammation enabling testing of new anti-inflammatory drugs in the model systems
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