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Background and Purpose: autogenous vein graft surgery is one of the important means of limb ischemic disease or coronary heart disease treatment, but postoperative graft restenosis, vascular occlusion affect its long-term efficacy. The study showed that the graft restenosis and vascular smooth muscle cell proliferation, but the exact mechanism has not been fully elucidated, which involves a series of cytokines, functional proteins and intracellular signal transduction pathway. Protein kinase-C (PKC) is a serine / threonine protein kinase associated with a variety of intracellular signal transduction process. Wherein the PKC-α is in one of its traditional isoenzymes, with a variety of cell proliferation, differentiation, but unclear whether to participate in the proliferation of smooth muscle of the vein graft. This study using autologous vein graft model change detection within the vein graft the PKC-αmRNA protein, to further clarify the relationship with the proliferation of vascular smooth muscle cells, then looking for a new prevention autogenous vein graft stenosis The method provides a theoretical basis. Method: 1, the establishment of the rabbit neck autologous vein graft model. 2, light microscopy intimal hyperplasia after transplantation pathological conditions. 3, observed after transplantation, blood vessels the PKC-αmRNA, protein expression changes dynamically. 3.1 Real-Time PCR assay vein graft PKC-αmRNA expression. 3.2 Western Blot assay vein graft PKC-α protein expression. 3.3 SP immunohistochemical staining the vein graft PKC-α protein expression. Results: 1 establishment of the rabbit neck autologous vein graft model. 1.1 during the experiment, the vascular anastomosis failure, two weeks after the surgical vein graft occlusion. The loss of the samples were found to make up. 1.2 naked eye: Adapted see animals incision healed well, no infection, mild adhesion of the vein graft and surrounding tissue, vein graft beat well, different degree of thickening of the wall of the vein graft luminal irregularity was arteriovenous performance. 2, light microscopy intimal hyperplasia after transplantation pathological conditions. Light microscope the normal sham group intimal endothelial monolayer cells, in a thin film, and is composed of 2 - layer 3 of vascular smooth muscle cells and few collagen. Days after transplantation visible intimal damage, inflammatory cell infiltration. Of 7 days after transplantation visible intimal hyperplasia, 14d and 28d, visible endometrial gradually thickening of smooth muscle cells, vascular lumen relatively smaller. 3, observed after transplantation, blood vessels the PKC-αmRNA, protein expression changes dynamically. 3.1 Real-Time PCR assay vein graft express PKC-αmRNA,: relative expression level of mRNA was significantly increased in the first three days after the transplant is about the control group (2.335 ± 0.059) times the difference between the two groups was statistically significance (P lt; 0. 01). With time, the amount of the expression is gradually reduced. 3.2 Western Blot assay vein graft PKC-α protein expression: PKC-α expression in 3 days after transplantation was significantly higher, and then gradually decline. PKC-α protein, 3.3 SP immunohistochemistry staining of vein graft: cytoplasmic expression most of the positive cells in thickened intima and intima, hyperplasia tissue expression of positive cells was significantly higher than that of non-proliferative part hyperplasia, pale-brown particles also increased significantly. Conclusion: Successful the rabbit neck autologous vein graft model; optical microscope to transplant intimal damage, hyperplasia, with time, the vessel lumen is relatively small; early vein graft, PKC- αmRNA its protein expression is elevated, then gradually decreased to normal, and its expression is targeted at the proliferation of smooth muscle cells. This suggests that PKC-α may be involved in the initial process of vascular smooth muscle proliferation signal transduction.
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