Relationshio pregenitor cells derived from bone marrow and graft intimal hyperplasia

基于LacZ小鼠颈动脉间置下静脉内膜增生的认识，我们制作了移植模型，但由于技术上的困难，我们不得不放弃。除此之外，我们继续用家兔制作接枝模型。（方法）将家兔随机分为两组；对照组饲喂市售兔粮，普伐他汀组和普伐他汀组分别饲喂普伐他汀钠和普伐他汀钠10mg/kg。在适当麻醉下行颈动脉介入-颈静脉逆行移植。在植入术后的每个时间点，在全身麻醉下摘取自体静脉。移植物的中间部分用于组织学和免疫组织化学研究。术后4周，两组均行静脉移植，镜下观察血管内膜增生。植入2周后，收集更多的细胞后进行PCNA抗体染色。为了计算该PCNA指数，在随机8个视野中计数PCNA阳性细胞和总细胞。为了评估细胞凋亡活性，我们采用TUNEL法检测原位细胞凋亡。将阳性细胞数除以计数细胞总数定义为移植物的TUNEL指数。此外，还购买了人脐静脉内皮细胞（HUVECs）和血管平滑肌细胞（VSMCs）。用1-μ m和30-μ m普伐他汀处理细胞24小时，用10%三氯乙酸收获。所得沉淀物用抗磷酸化mypt -1、抗mbs抗体、抗rhoa和抗enos /NOS III型进行免疫印迹。（结果）正常胆固醇血症家兔口服亲水他汀、普伐他汀和匹伐他汀可抑制血管移植4周后血管增生，抑制血管移植2周后血管内膜细胞增殖和凋亡。此外，我们发现普伐他汀抑制rho激酶活性，加速内皮细胞，而不抑制血管平滑肌细胞的rho激酶活性。（结论）亲水性他汀类药物可通过内皮细胞选择性抑制rho激酶抑制晚期静脉移植衰竭。此外，这些结果强烈支持临床使用亲水性他汀类药物来预防搭桥术后晚期移植物衰竭。少

Based on that intimal hyperplasia is recognized in the interposed inferior vena in the carotid arteries of LacZ mouse, graft model was made, However, it was very difficult to performe technically and we could not help giving it up. Otherwise we kept on making graft model with the use of rabbit. (Methods) Rabibits were randomly divided into two groups ; the control group that was fed commercial rabbit chow, and the pravastatin or pivatastatin group that received the same chow with 10mg/kg pravastatin or pivatastatin sodium. Carotid interposition-reversed juglar vein grafts were performedunder appropriate anesthesia. At each time point after implantation of the autologous vein grafts, the vein grafts were harvested under general anesthesia. The middle portion of the graft was used for histological and immunohistochemical studies. Four weeks after operation, vein grafts in both groups were harvested and intimal hyperplasia were elucidated macroscopically. After 2 weeks of implantation, gr … More aft were harvested and stained with PCNA antibodies. To calculate this PCNA index, PCNA positive cells and total cells were ciunted at random 8 fields. In order to evaluate apoptosis activity, we used the TUNEL method to detect in situ apoptosis. The number of positive cells divided by the total number of cells counted was defined as the TUNEL index of the graft. Additionally human umbilical vein endothelial cells(HUVECs) and vascular smooth muscle cells(VSMCs) was purchased. The cells were treated with 1-μ Mand 30-μM pravastatin for 24 hours and harvested with 10% trichloroacetic acid. The resulting precipitates were subjected to immunoblotting with anti-phospho-MYPT-1,anti-MBS antibody, anti-RhoA, and and anti-eNOS/NOS type III. (Results) We demonstrated that oral administration of the hydrophikic statin, pravastatin and pitavastatin, to normocholesterolemic rabbits inhibited hyperplasia of the vein grafts 4 weeks after implantation, and suppressed cell proliferation and apoptosis in the neointima 2 weeks after implantation. In addition, we found that pravastatin inhibited Rho-kinase activity and accerated endothelial cells, while it did not inhibit Rho-kinase activity in vascular smooth muscle cells. (Conclusion) Based on our findings hydrophilic statin can suppress late vein graft failure through endothelial cell-selective inhibition of Rho-kinase. Furthermore, these results strongly support the clinical use of hydrophilic statins to prevent late graft failure after bypass grafting. Less