Figure 1: Expression of TSG-6 in Human Vascular Cells and its Effects on Inflammatory Response and Proliferation in Human ECs
2019-06-11T07:02:50Z (GMT) by
(A) Aliquots of 40 μg of cellular protein from human monocytes, HMDMs, HASMCs, and HUVECs were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subjected to immunoblotting analysis for human TSG-6 and β-actin. Immunoblots are representative of 2 independent experiments. (B) Proliferation of EA.hy926 ECs was determined by WST-8 assay after 48-h incubation in DMEM containing 2 concentrations of d-glucose (Glc) (1.0 or 4.5 mg/ml) with the indicated concentrations of TSG-6. EA.hy926 ECs cultured in 4.5 mg/ml d-glucose–conditioned medium were stained by hematoxylin eosin. Glc 1.0 mg/ml (blue): n = 4; *p < 0.005; †p < 0.0005; ‡p < 0.0001 vs. 0 ng/ml of TSG-6. Glc 4.5 mg/ml (red): n = 5; #p < 0.01 vs. 0 ng/ml of TSG-6. (C) HUVECs were pre-treated with or without TSG-6 (300 ng/ml) for 30 min and then incubated with or without LPS (1 μg/ml) for 2 h. The levels of IL-6, TNF-α, MCP-1, ICAM-1, VCAM-1, E-selectin, and GADPH mRNA expression were measured by reverse transcription polymerase chain reaction. The graph shows the densitometric data of each molecule after normalization relative to GADPH. n = 3 to 4; §p = 0.028; ¶p < 0.005 vs. control; ＠p < 0.01; &p = 0.028 vs. LPS. All values are presented as mean ± SEM. EC = endothelial cells; GAPDH = glyceraldehyde-3-dehydrogenase; Glc = d-glucose; HASMC = human aortic smooth muscle cell; HMDM = human monocyte-derived macrophage; HUVEC = human umbilical vein endothelial cell; ICAM = intercellular adhesion molecule; IL = interleukin; LPS = lipopolysaccharide; MCP = monocyte chemotactic protein; mRNA = messenger ribonucleic acid; TNF = tumor necrosis factor; TSG = tumor necrosis factor–stimulated gene; VCAM = vascular adhesion molecule.