| Purpose:Through establishing the animal models that are displaced their pulmonary artery, thoracic aorta and inferior vena cava artificial blood vessels, to compare the morphology of pulmonary and systemic circulation of artificial vascular intima, the expression ultrastructure and TPAmRNA and protein, so as to explore the Significance expression of TPA in vascular endothelial cells in different blood flow environment.Methods:There are twenty-seven pigs, according to the principles of randomized into three groups:Group of replacing pulmonary artery (Pulmonary Artery Reconstruction via e-PTFE Artificial Blood Vessels), n=9; Group of replacing thoracic aorta(Thoracic Aorta Reconstruction via e-PTFE Artificial Blood Vessels), n=9; Group of replacing inferior vena cava(Inferior Vena Cava Reconstruction via e-PTFE Artificial Blood Vessels), n=9; Each vessel group was further divided into 2 week of March, June, three time point subgroups, there are three pigs at each time point group; The establishment of animal model of artificial vascular replacement after positive postoperative nursing by professional breeders of conventional farming. After postoperative 2 weeks, three months, six months respectively exam by color Doppler ultrasound and take out artificial vascular, measure and replace the stenosis degree of vascular, with or without tarombokinesis and so on. Take out the e-PTFE artificial vascular and analysis the degree of endothelialization of artificial vascular and the ultrastructure of endothelial cell based on naked eye observation, optical microscopy, scanning electron microscopy under different environment. Test the TPA expression of artificial vascular endothelial cells in each group by immunohistochemical method; Test the TPA-Protein level of artificial vascular endothelial cells in each group by Western Blot. Test the TPAmRNA level of artificial vascular endothelial cells in each group by qRT-PCR. The detection results were statistically analysis.Results:First, all experimental pigs survived, in good condition, animal models were successfully established.Second, Vascular ultrasound results showed pulmonary artery group, thoracic aortic group heart function were normal, don’t show any signals of tarombokinesis; inferior vena cava group has obvious tarombokinesis, and inferior vena cava widened, part of the visible image of ascites.Third, naked eye observation:pulmonary artery group, thoracic aortic group both has obvious tarombokinesis by taking out the e-PTFE artificial vascular about two weeks, but the caliber’s stenosis of artificial vascular does not exceed 50%; After postoperative three months, six months each artificial vascular has not, obvious tarombokinesis; However, inferior vena cava group has tarombokinesis after two weeks, meanwhile, the caliber’s stenosis of artificial vascular exceed 50%, it show adhering tarombokinesis on caliber in three months and six months, but less in six months.Fourth, morphological observation, HE staining:the growth of implanted artificial vascular ECs is normal, no abnormal cell growth or tumor cell growth. Scanning electron microscope:The thoracic aorta can be seen in the newborn endothelial cells are spindle, in pulmonary artery endothelial cells visible newborn are long oval, spindle shaped, and the new low shear stress in the inferior vena cava in endothelial cells are round, cobblestone, arterial endothelial cells arranged more tightly and arranged in vein endothelial cells relatively loose. Immunohistochemistry: It’s visible with DAPI dye staining of ECS cells show blue oval, polygon area.594 dye dyed red area is for TPA protein, it can be seen it express in the cell cytoplasm. At time goes on, there is a significantly decreasing trend of TPA expression in each group.Fifth, Molecular biological detection results, qRT-PCR tests the TPAmRNA expression level:pulmonary artery group versus thoracic aortic group don’t exist statistically significance (P>0.05); Inferior vena cava group versus pulmonary artery group, inferior vena cava group versus thoracic aortic group exist statistically significance (P<0.05); Test the TPA-Protein expression level by Western Blot: pulmonary artery group versus thoracic aortic group don’t exist statistically significance (P>0.05); After postoperative 2 weeks, three months, inferior vena cava group versus pulmonary artery group, inferior vena cava group versus thoracic aortic group exist statistically significance (P<0.05); After postoperative six months, inferior vena cava group versus pulmonary artery group, inferior vena cava group versus thoracic aortic group don’t exist statistically significance (P>0.05).Conclusion:First, implanted e-PTFE artificial vascular in the animal body caused caliber’s stenosis with thrombosis, influence the vascular patency. Implanted e-PTFE artificial vascular in pulmonary artery group and thoracic aortic group are close to patency, but the patency of implanted e-PTFE artificial vascular in inferior vena cava group are poor than other group.Second, the growthform of implanting e-PTFE artificial vascular in pulmonary artery and thoracic aortic of ECs is similar, the direction of the long axis and the blood flow in the same direction. However, the growthform of implanting e-PTFE artificial vascular in inferior vena cava exist obvious difference to vascular ECs, the direction of cell growth was not obvious.Third, the TPA level of implanting e-PTFE artificial vascular in pulmonary artery and thoracic aortic of ECs secret higher than implanting e-PTFE artificial vascular in inferior vena cava. After two weeks, three months,there are no difference between pulmonary artery and thoracic aortic in the secreting of VECs level and TPA level, both of two groups’ TPA level secrets higher than inferior vena cava group, after postoperative six months, the TPA expression level of each group is closed. |
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