Hepatocyte Growth Factor Experimental Research Of Chronic Deep Vein Thrombosis Organization Recanalization

Objective:To investigate the role of HGF in promoting organization,angiogenesis,recanalization of thrombus by constructing adenovirus hepatocyte growth factor(HGF)recombinants and its use in the model of deep vein thrombosis of the lower limbs of house dogs.Methods:1?The peripheral blood of dogs was isolated and the white blood cells were isolated.Total RNA was extracted with Trizol reagent and reverse transcribed into cDNA.The target gene HGF was amplified by PCR and cloned into adenovirus vector.The recombinant virus was packed in 293 A cells,which was further purified and concentrated by ultracentrifuge and dialysis.2?A total of 18 normal house dogs(15Kg-18kg)were selected(Male or female).By using the technique of interventional Seldinger puncture technique combined with traditional open surgery,animal model of iliac vein compression and chronic deep vein thrombosis was simulated.18 dogs with chronic deep vein thrombosis were randomly divided into three groups: control group,anticoagulation group and treatment group,6 dogs in each group.Dogs were injected with normal saline,low molecular weight heparin,Adenovirus of HGF respectively.28 days after feeding,dogs were observed by DSA to detect the thrombosis andcollateral circulation.The expression of HGF mRNA in the thrombus was detected by real-time fluorescence quantitative PCR.The expression of HGF protein was detected by western blot.Immunohistochemical staining to vWF was performed to measure the density of angiogenesis.Result:1? Adenovirus with the target gene hepatocyte growth factor was successfully constructed.The green fluorescence could be observed under the microscope.The titer of recombinant virus was about 1.0 × 108/ml.2?Dog model of deep vein thrombosis was established.DSA angiography confirmed deep venous thrombosis of lower limbs.After treatment,the number of collateral vessels were significantly higher in the treatment group than in the control group and the anticoagulation group(p <0.05)detected by DSA.The expression of HGF mRNA of thrombus in the treatment group was significantly higher than that in the control group(P <0.05).WB showed that the expression of HGF protein in the treatment group was higher than that in the control group and the anticoagulation group(p <0.05).The number of capillaries measured by vWF density in the treatment group was significantly higher than that in the control group and the anticoagulation group(p <0.05).The success of the hepatocyte growth factor gene into the adenovirus,with the green fluorescent drops to 1.0 x 108 hepatocyte growth factor of the adenovirus recombinant.Conclusion:1? hepatocyte growth factor can obviously promote angiogenesis in chronic thrombus formation.2?hepatocyte growth factor can obviously accelerate the chronic thrombosis machine and recanalization.