P27 < Sup > Kip1 < / Sup > Impact On Canine Carotid Stent Restenosis And Shu, The Study Of The Intervention

Part I Establish the models of carotid atherosclerotic stenosis in BeaglesObjective:To explore whether the models of carotid atherosclerotic stenosis in Beagles can be established by the way of balloon injury and nitrogen drying of endothelial cells in their carotid arteries with high-fat dietMethods:19healthy beagles in male were randomly divided into2groups, group A (sham group, n=4) and group B (atherosclerotic group n=8). Right common carotid artery was only separated in group A,not injuried; group B was injuried by the way of balloon injury and nitrogen drying of endothelial cells in their carotid arteries, both of them supported by high-fat diet. Vascular ultrasound was used to learn the basic situation of the common carotid artery before modeling, which was used to observed the carotid artery stenosis and atherosclerotic plaque formation two months after modeling with DSA. P<0.05represent significant statistically differences by the software of SPSS17.0.Results:Carotid intima of group A is smooth, which show by results of the vascular ultrasound, without stenosis and plaque formation; Carotid arteries of group B can be seen varying of degrees of stenosis and plaque formation. DSA results show contrast group B to group A, group B was less than group A in the minimum diameter (2.352±0.839vs3.080±0.13, P=0.010), and group B is much larger than group A in stenosis rate(34.240±18.004VS4.727±2.062, P=0.000).Conclusion:The good model of carotid atherosclerotic stenosis in Beagles can be established by the way of balloon injury and nitrogen drying of endothelial cells in their carotid arteries with high-fat diet Part II: A researchon the effect of p27KIP1in in-stent restenosis of beagles carotid with intervention of FasudilObjective:To Observe the relationship between Rho kinase activity after Carotid artery stenting and the expression change of P27KIP1to neointimal formation, and to learn the effect of prophylaxis and treatment in the process of in-stent restenosis by inhibiting Rho kinase activity.Methods:The4beagles still was group A (sham operation group, n=4) as previous, and the8previous beagles with modeling of the carotid artery atherosclerosis were randomly divided into group B (group of bare stent) and group C (group of bare metal stent+fasudil). Bare metal stents were placed into the right common carotid artery of group B and group C. Fasudil was given to group C two hours after intervention (intravenous infusion,3mg/kg/time,2times/day,seven days totally). DSA and vascular ultrasound were examined to observe the situation of the carotid restenosis two months later, then remove the right common carotid artery. HE and elastic fiber stainings observed pathomorphological situation; Western blot assayed the level of expression of P-MYPT-1, P27KIP1. P<0.05was considered significant statistically differences by SPSS17.0for analysis of One-way ANOVA.Results:1. Vascular ultrasound showed no thrombosis in the lumens two months later after stenting. DSA results showed the lumens of group A were smooth without stenosis; the minimum diameter of group B was less than group C, whose differences has statistically significances(P=0.000); group B is higher than group C in stenosis rate, the differences has statistically significances(P=0.000).2. HE and elastic fiber stainings showed no intima hyperplasia in group A, but was significant in group B and C; the neointimal thickness, area and stenosis rate in group B were higher than group C, the differences has statistically significances(P=0.000).3. Western blot showed weak expression of P-MYPT-1in beagles carotid of group A. but increased in group B and C, especially higher in group B, the differences has statistically significances (group B VS A, p=0.000, group C VS A p=0.045); P27KIP1had a higher expression in beagles carotid of group A, but reduced in group B and C, especially lower in group B, the differences has statistically significances (group B VS A, p=0.000, group C VS A p=0.045).4. Immunohistochemical staining of a-SMA showed a large number of a-SMA expressed in media of all three groups. Group A appeared on expression of a-SMA-positive outside of the elastic intima, but opposite in the new intima of groups B and C; the mean optical density value of a-SMA in group B is higher than in group C, the difference has statistically significances(P=0.001). 5. Pearson correlation coefficient analysis showed:P-MYPT-1and P27KIP1,a-SMA and Intimal area(IA) was positively correlated(r=-0.704,0.934,0.934);P27KIP1and a-SMA,IM was significant correlation (r=-0.740,-0.621).Conclusion:1. Rho kinase activity and neointimal hyperplasia increase obviously, which indicates that Rho kinase participate the process of in-stent restenosis after Carotid artery stenting.2. Fasudil may reduce the expression of p27KIP1by inhibiting Rho kinase activity markedly, indicating that Fasudil may reduce neointimal hyperplasia by up-regulating the expression of p27KIP1, so as to prevent the formation of the ISR.