Endothelial HMEC-1 cells incubated with pro-inflammatory cytokine TNF-α for 6 and

Endothelial HMEC-1 cells incubated with pro-inflammatory cytokine TNF-α for 6 and 24?hours were studied being a model of inflammation using Raman imaging. being a hallmark of inflamed cells. The statistical analysis showed that the number of lipid body 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. Basic knowledge about the subcellular changes that occur inside the cell body during numerous cellular events and under disease conditions is extremely important for understanding of the mechanisms of pathology development increasing the chance of successful diagnostics and treatment. Confocal Raman microscopy is usually a powerful tool to study cellular model systems due to several reasons: 1. the possibility to obtain comprehensive information about the biochemical composition of the sample 2 the submicrometer Otamixaban spatial resolution providing information about the changes in the biochemical composition and their location at the subcellular level and 3 the sensitivity and structural specificity in label-free detection of alterations occurring in organelles and main cellular compartments. All pointed out advantages make Raman microscopy a particularly convenient tool to develop understanding of cellular processes in diseases. One of many potential branches of medicine in which Raman microscopy can be used successfully is usually cardiology. The growing evidence indicates that some cardiovascular events and way of life diseases i.e. atherosclerosis diabetes and hypertension begin with endothelial dysfunction1 2 3 4 5 and thereby endothelial cell cultures are convenient models to study pathology development in the circulatory system. However only few reports have been previously published regarding human endothelial cells lines cultures studied by means of Raman microscopy6 7 Otamixaban 8 9 10 11 So far it has been applied to characterize the chemical and structural changes related to early apoptosis invoked by exposure to numerous agents6 formation of lipid droplets7 8 and its Rabbit Polyclonal to SLC10A7. role as a cargo in intercellular communication9 or to monitor the intracellular pH10 and accumulation of bioactive drugs i.e. anthracyclines in studies on mechanisms of endothelial toxicity11. The endothelium is usually a highly specialized unicellular layer of cells lining the blood and lymph vessels the natural gatekeeper between the blood and the vascular wall which is responsible for a wide variety of crucial processes controlling vascular functions1 12 One of the main actions of endothelial cells is usually associated with triggering of innate and acquired immune response after activation by e.g. pro-inflammatory cytokines hypoxia or metabolic stress in which they produce and release cytokines and growth factors sending signals to leukocytes13 14 15 The action of pathological brokers and traditional risk factors including among others aging smoking and hypercholesterolemia provokes endothelial dysfunction a process characterized by diminished production or availability of nitric oxide2 16 Moreover it is also suggested that the early step of endothelial dysfunction manifests itself in developing of inflammatory says16. Inflammation in general is intended to limit invasion and damage after the injury a process that is essential for the survival of organisms. It involves both adaptive and innate defense systems concerning general and specialized type Otamixaban of protection against foreign pathogens respectively. In the mobile range endothelial cells react to pro-inflammatory elements via a number of different systems. Activation may appear through particular membrane receptors i.e. the tumor necrosis aspect receptor 1 (TNF-R1) by binding of its ligand tumor necrosis aspect alpha (TNF-α) the interleukin 1 receptor type I (IL-1R1) after arousal by interleukin 1 (IL-1) or toll-like receptor (TLR) when endothelial cells face endotoxins13 15 All talked about right here inflammatory mediators switch on endothelial cells through the traditional nuclear aspect kappa-light-chain-enhancer of turned on B cells Otamixaban (NF-κB) pathway. NF-κB induces activation of varied elements.