Laboratory Research On The Correlation Between PKC-α And Angiogenic Remodeling After Autogenous Vein Transplantation

Background and Objectives:The transplantation of autogenous vein is the most important method to cure mutilation diseases and coronary heart diseases. However the transplant may narrow down, so it will lead to vascular occlusion which affects it's long-term healing effect. The narrowing down of the transplant is related to the hyperplasia of vascular smooth muscle cells, but its specific process has not been clearly illustrated. What has been known is that it refers to a series of cytokines,functional proteins,intracellular signal transduction pathwaysEtc. PKC is one of serine / threonine protein kinase which is related to varies of intracellular signal transduction. PKC-αis one of the traditional isozyme and is related to varies of cell proliferation, differentiation, yet it has not been proved if it's relevant to Smooth muscle cell proliferation of the Vein graft. This research intends to adopt the model of rabbit autogenous transplantation of veins to further illustrate its relation to smooth muscle cell proliferation by testing the changes of PKC-αmRNA and protein of the vein graft and it will provide the theoretic foundation for the new treatment to prevent Autogenous vein graft restenosis.Methods:1,Buildind of the model of rabbit neck autologous vein graft.2,Observation of the pathology of intimal hyperplasia after transplantation by light microscope.3,Dynamic observation of the expression of PKC-αmRNA, protein expression. after transplantation 3.1 Detection of PKC-αmRNA expression. by Real-Time PCR3.2 Detection of PKC-αprotein expression. by Western Blot3.3 Detection of PKC-αprotein expression. by SP immunohistochemical stainingResults:1,Building of the model of rabbit neck autologous vein graft.1.1 Situations in the experiments: one experiment suffered from the failure of vascular anastomosis, and the other experienced vein blockage two weeks after the experiment.1.2 Naked eye: the animals when derived wound healing well, no infection, mild vein graft and the surrounding tissue adhesion, good vein graft pulsation, to varying degrees of vein graft wall thickening, luminal irregularity, was the performance of venous arterialization.2,Observation of the pathology of intimal hyperplasia after transplantation by light microscope.Light microscope, the sham group was normal intimal endothelial cell monolayer, the film is thin, from 2 - 3 layers of collagen in vascular smooth muscle cells and a small amount of the composition. 3d after transplantation, intimal visible damage, inflammatory cell infiltration. Significantly after transplantation, intimal hyperplasia can be seen 7d, 14d and 28d at the time, showing the gradual thickening of the intima, with more smooth muscle cells, blood vessel lumen is relatively small.3,Dynamic observation of the expression of PKC-αmRNA, protein expression. after transplantation3.1 Detection of PKC-αmRNA expression. by Real-Time PCR: in the first 3 days after transplantation, mRNA expression significantly increased the relative about the control group (2.335±0.059) times the difference between the 2 groups was statistically Significance (P <0. 01). With time, the expression level gradually decreased.3.2 Detection of PKC-αprotein expression. by Western Blot . Expression of PKC-αon day 3 after transplantation was significantly higher, and then gradually decreased.3.3 Detection of PKC-αprotein expression. by SP immunohistochemical staining Positive cells mostly in the medial and intimal thickening, proliferation of the number of positive cells in tissues was significantly higher than non-proliferation section, and the brown granules in the cytoplasm of proliferating cells was significantly increased.Conclusions:Established a rabbit model of vein graft from the neck successfully observed under light microscope after transplantation, intimal damage, hyperplasia, relatively smaller lumen with time, in the vein of early after transplantation, PKC-αmRNA and protein expression increased, then decreased gradually normal, and its expression localized in the proliferation of smooth muscle cells. This indicates that PKC-αmay be involved in the initial process of vascular smooth muscle signal transduction.