Tissue Factor Pathway Inhibitor Effects Tissue Factor In Early Stage Atherosclerosis

Objective:The objective of this study is to evaluate whether to add the low dose of tissue factor pathway inhibitor(TFPI) long time decreases tissue factor(TF) to ApoE-/- mice by plasmid VRnew in the early stage of atherosclerosis.Methods:Feed three 8-week-old female ApoE-/- mice with west diet.Transfect mice with 60ug VRnew-TFPI plasmids at their muscle of posterior limbs after 2 weeks,then take mice's blood at 4th,14th and 20th day after transfaction,Add natrium citricum solution as 1:9(v:v) for anticoagulation.Measure the concentration of blood human TFPI by ELISA kit.Feed 14 6-week-old female ApoE-/- mice through the construction with west diet,According to weight of mice in 8-week to divide into 2 groups.Inject 30ul plasmid(concentration lug/ul) into muscle of every posterior limb of mice then 150V/cm square pulse shock 8 times after 2 weeks. VRnew—TFPI group(7 mice) transfects VRnew-TFPI plasmid and VRnew group(7 mice) transfects VRnew empty plasmid.Repeat every two weeks,total 6 times. 20-week-old mice are sacrificed and take aorta.Make paraffin sections.Stain TF antigens by immunohistochemistry.Employ with Image-Pro^? Plus V6.0 to deal with the result of immunohistochemistry,read optical density and area,count average density.Analyze the result.Results:Measure the plasma concentration of human TFPI by ELISA,4^th day after transfect plasmid 2.274±0.594 ng / ml;14^th day 1.676±0.069ng/ml;20^th day 0.486±0.3325ng/ml.Divide 14 8-week-old mice into two groups according to weight at that time,the average weight of VRnew—TFPI group is 18.34±1.90g and the average weight of the VRnew group is 18.85±1.39s.P＞0.05.We can observe white atherosclerotic plaque in the aorta after the mice were sacrificed,then observe plaque under microscope after HE staining.Immunohistochemistry method marks TF antigen on the aorta,Image-Pro^? Plus V6.0 software reads optical density and area,calculate average optical density value,VRnew—TFPI group value is 0.23±0.08 and VRnew group value is 0.34±0.10,P＜0.05.Conclusions:Given long-term low-dose TFPI,tissue factor in mice's aorta will be decreased in the early stage of atherosclerosis.