Sinomenine inhibited TNF-α induced NF-κB p65 level in human umbilical vein endothelial cells nucleus

Objective To investigate the effect of sinomenine(SIN)on TNF-α induced NF-κB p65 level in human umbilical vein endothelial cells(HUVECs)nucleus.Methods HUVECs were isolated from freshly collected umbilical cords. Positive controls samples were stimulated with TNF-α,omitting SIN. Negative controls samples were treated in the same way,omitting TNF-α and SIN. Experiment samples were co-cultured with TNF-α and SIN at different concentrations (0.25,0.5,and 1.0 mol/L),or TNF-α and dexamethasone(Dex) at concentration of 1.0×10-6 mol/L.Cell nuclear extracts were prepared according to extraction procedure of Nuclear Extraction Kit .NF-κB p65 in cells nuclear extracts was detected by ELISA.Results The protein concentration of cell nuclear extracts samples were from 0.16 g/L to 0.77 g/L. NF-κB p65 levels in cell nucleus in the groups of SIN (at concentration of 0.25 mol/L to 1.0 mol/L) and Dex (at concentration of 1.0×10-6 mol/L) decreased compared with the group of positive control.Conclusion SIN and Dex inhibit NF-κB p65 level of cell nucleus in HUVECs stimulated by TNF-α in vitro. Key words: Sinomenine; Umbilical vein; Endothelial cells; Nucleusfactor-κB; Tumor necrosisfactor-α