Effect Of Adiponectin On Expression Of Chemotatic Factor And The Level Of MRNA Of Adhesion Factor In Umbilical Vein Endothelial Cells

ObjectiveAdiponectin is a recently discovered factor,which specificly secreted by adipose tissue, with increased insulin sensitivity, inflammation, anti-atherosclerosis and other effects. The levels of adiponectin in vivo is closely related to the process of many inflammatory diseases, especially coronary atherosclerotic heart disease. However, the mechanism of adiponectin against atherosclerosis needs further study.To investigate the effect of adiponectin on expression of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) and the influence of adiponectin on the mRNA level of adhesion factor in umbilical vein endothelial cells induced by tumor necrosis factor-α(TNF-α).Methods1,Human umbilical vein endothelial cells (HUVECs) were isolated from umbilical vein, which were then indentified by the detection of endothelial cell-specific factorⅧ-related antigen with the use of labeled streptomycin avidin-biotin method (LSAB method) and fluorescence staining.2,HUVECs were cultured in vitro and divided into control group, TNF-αstimulated group (5, 10, 20, 50μg/L TNF-α) and adiponectin intervention group (5, 30, 50mg/L adiponectin +20μg/L TNF-α).3,RT-PCR and ELISA were used to detect IL-8 and MCP-1 expression after treatment.4,Last compared with TNF-αstimulation group in the mRNA levelsof vascular adhesionmolecule-1(VCAM-1), intracellular adhesion molecule-1(ICAM-1) and E-selectin by RT-PCR.5,With SPSS13.0 statiatical software,carry out statiatics analysis ,Significance was set at P<0.05.Results1,Fluorescence staining showed thatⅧfactor related antigen is expressed in the cytoplasm, which confirmed that the cultured cells is the vascular endothelial cells.2,The expressions of IL-8 and MCP-1 were markedly increased in HUVECs after stimulation of TNF-α(P<0.01), also it is dose-dependent. There was no significant difference regarding IL-8 and MCP-1 expression between 20μg/L TNF-αstimulated group and 5mg/L adiponectin intervented group. However, IL-8 and MCP-1 expression were significantly decreased in 30 mg/L and 50mg/L adiponectin intervented group compared with 20μg/L TNF-αstimulated group (P<0.01).3,TNF-αstimulation group increased mRNA levels of VCAM-1, ICAM-1 and E-selectin. Compared with TNF-αstimulation group,the mRNA levels of VCAM-1, ICAM-1 and E-selectin. in adiponectin and TNF-αassociated stimulation group were decreased.ConclusionAdiponectin has protective actions against the initiation and progression of atherosclerosis via repressing TNFα-induced IL-8 and MCP-1 and adhesion factor (such as VCAM-1, ICAM-1 and E-selectin)expression.