A Study Of Relationship Between DVT And MCP-1Expression In Blood

Objectives1. To create mice DVT model, observe animal thrombus formation in different time points, MCP-1expression is detected in the blood of mice; to explored the relationship of MCP-1and DVT in mice.2. To detect MCP-1expression level at different stage of DVT in human blood,,, to explore the relationship of MCP-1and DVT in human blood.3. Through the study of gene expression and found that MCP-1gene upregulation. To combined genetic information analysis techniques, we found endothelial cell injury, ERK signaling pathway activation leads MCP-1gene expression, platelet activation, coagulation enhanced further elucidate the molecular mechanism of thrombosis to be more targeted, for the discovery of high sensitivity, the new quantitative diagnosis of DVT and provide experimental evidence of molecular targets.Materials and Methods:Part1To create mice DVT model, and detect MCP-1expression level at different time points in DVT mice.1.104Kunming mice were fed for3to5days before the experiment. Eight normal mice were selected random as control group. The remaining96were divided into model group and sham operation group.48randomly selected as the model, ligation of the inferior vena cava by2h,4h,6h,8h,12h,24h time point chambers of the heart blood, take1.0cm from the ligature of the inferior vena cava and the contents of4%paraformaldehyde fixed evacuation paraffin sections at each time point bit8mice. And take the remaining mice sham-operated group, layer cut mouse skin, subcutaneous tissue, layer by layer to close the abdominal cavity, the separation of the inferior vena cava,2h,4h,6h,8h,12h,24h time point chambers of the heart blood each time point8.2. Naked eye generally observed thrombotic conditions and standards of judgment: modeling of the inferior vena cava color, degree of swelling, the degree of local tissue reaction observed after the abdominal cavity was opened; anatomy drawn under the microscope after the change of the inferior vena cava, the judge thrombosis3. Mice mining HE staining and sliced, observed at different time points adopted by the Organization of the lumen of the vein thrombosis.4. PCR was detected mice of each group blood cells in blood total RNA, the application Oligo6.69primer design software, provided for a semi-quantitative PCR reverse primer, each group of mice RNA is reverse transcribed into cDNA, and GAPDH as an internal reference, using Trizol extraction MCP-1expression Cover.Results1. The mouse model is complete after2h naked eye than the control group was not obvious. The4h group no obvious thrombosis. Mice after modeling6h vein wall incomplete thrombus formation; modeling8h vein wall complete thrombus formation, the inferior vena cava coronal plane can be seen in the venous lumen thrombosis contains red patchy distribution of red blood cells, lighter in color pink like platelets, scattered between the distribution of white blood cells, thromboembolic and vein wall is more closely connected. The12h and24h group of thrombus cracks appeared close to the vessel wall, a large number of leukocyte infiltration, close to the central location of the venous lumen thrombosis platelet lighter color around a small amount of white blood cells.2. We explored Pathway bioinformatics to analysis the process of thrombus formation expression of MCP-1, and endothelial cell injury, monocytes, platelet aggregation, blood coagulation.PART2PCR detection of human trauma group thrombosis do not form a group, the blood cells of the thrombosis group and the normal group of MCP-1changes1. Based on clinical observation, Color Doppler Ultrasound, venography combination of inspection, the experiment was divided into three groups; normal control group (A group), trauma group thrombosis do not form a group (B), thrombosis group (C group). My suffering, including spinal surgery, for knee, hip, lower limb fracture trauma thrombosis group and do not form a group within blood. Each each patient blood5ml, After centrifugation, the plasma and blood cell samples, blood total RNA extracted by Trizol, the specimens by PCR quantitative detection the MCP-1of blood.Result1. Using Trizol extraction of total RNA of blood the application Oligo6.69primer design software provided a semi-quantitative reverse transcriptase PCR primers, each of the groups of human blood RNA is reverse transcribed into cDNA, and GAPDH as an internal reference, ran Matches electrophoretic bands.2. To create mice DVT model, modeling animal thrombus formation in a different time point focus state, while Follow the aggregation of mononuclear cells and platelets in thrombosis. MCP-1expression is detected in the blood of mice; explore the relationship of MCP-1in the blood of mice with DVT.Conclusion1. Mouse model of MCP-1mRNA expression amount and modeling mouse biopsy thrombosis match, MCP-1and thrombosis is closely related to.2. MCP-1was highly expressed in the blood of patients with clinical DVT, is expected to become a clinical deep vein thrombosis predictive diagnostic markers.