Effects Of TNF-α On Tissue Factor Pathway Inhibitor-2 Expression In Human Umbilical Vein Endothelial Cells

BackgroundMore and more evidence indicates that inflammation plays a very important role in the occurrence, development and prognosis of coronary heart disease. The relationship between a variety of inflammation factors and coronary heart disease is a hot research field. Revealing the relation has important clinical significance. Many kinds of inflammation factors (such as C response protein, tumor necrosis factor-alpha, interleukin-6, etc) have varying increased degree of synthesis in coronary heart disease. Tumor necrosis factor–αcauses inflammation and damage by increasing the procoagulant activity in endothelial cells, promoting the pathological damage and functional changes in endothelial cells mediated by activated leukocyte, which resulted in the plaque unstability and plaque rupture, then further the occurrence of acute coronary syndrome. Thrombosis in the blood vessel is the common pathological process in many diseases, which involves kinetic equilibrium among blood coagulation, anticoagulation and fibronolysis. Tissue factor pathway has the vital significance in the physiology hematischesis and the pathological thrombosis. Tissue factor pathway inhibitor is the main anticoagulant factor in vivo, which contains tissue factor pathway inhibitor-1 (TFPI-1) and tissue factor pathway inhibitor-2 (TFPI-2). This experiment is to investigate the effect of inflammation factor TNF-αon tissue factor pathway inhibitor 2 expression and mechanism in human umbilical vein endothelial cells.ObjectiveTo investigate the effect of TNF-α(20ng/mL), ERK blocker PD (10umol/L), NF-κB inhibitor prolidine dhiocarbamate (PDTC) (50umol/L) on TFPI-2 expression in human umbilical vein endothelial cells, then to explore the celluar mechanism.Methods(1) To culture human umbilical vein endothelial cells: primary human umbilical vein endothelial cells and low serum medium are purchased from the United States Cascade Biologics company. Culture the cell after resuscitation in 37℃, 5%CO2 incubator, through passage and freezing, uses the 3-4 generation cell.(2) HUVECs within 3-4 passages were divided into culture medium only group, TNF-αgroup, PD group, PDTC group. Protein levels of TFPI-2 were measured by western blot and TFPI-2 mRNA levels were assessed by quantitative real time RT-PCR.Results(1) TNF-αsignificantly increased the expression of TFPI-2 protein (P<0.01), ERK blocker PD and NF-κB inhibitor PDTC reduced the quantity of TFPI-2(P <0.05).(2) TNF-αsignificantly increased the expression of TFPI-2 mRNA (P<0.01), ERK blocker PD and NF-κB inhibitor PDTC inhibited the synthesis of TFPI-2 mRNA (P <0.05).Conclusion(1) TNF-αsignificantly increased the expression of TFPI-2, which is beneficial to prevent thrombosis development during inflammation(2) ERK blocker PD and NF-κB inhibitor PDTC inhibited the expression of TFPI-2, which illustrate mitogen-activated protein kinase signal transduction pathway (ERK) and NF-κB transcription factor has a positive regulation on TFPI-2 expression respectively.(3) Activating ERK pathway and NF-κB may be one possible pathway that TNF-αup-regulate the expression of TFPI-2 .