tPA基因局部定位转染抑制兔动脉损伤后内膜增生的研究

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 目的: 观察局部转染组织型纤溶酶原激活物 (tPA)基因, 对手术损伤兔右髂外动脉后内膜增生的影响并探讨可能的机制。方法: 采用显微外科手术方法, 建立兔右髂外动脉损伤模型。将 105只新西兰大白兔随机分为 3组, 每组 35只。A组为生理盐水对照组, B组为脂质体介导的pBudCE4. 1转染组, C组为脂质体介导的pBudCE4. 1 /tPA转染组。用微注射器将各种转染液注入损伤的血管壁, 每组按实验终点(术后 2、3、7、14、28d)再分为 5个亚组, 每个亚组 7只兔。于术后各实验终点, 取损伤段的血管用于病理学检查、电镜观察、RT PCR和免疫组化染色检查。结果: 与A组和B组相比较, 术后各时间点C组血管内膜的厚度、内膜的面积和血管腔的狭窄率均显著减小 (P<0. 01)。术后 28d时, C组血管腔的狭窄率比A组和B组分别降低了 51. 5%和54. 2%。术后各时间点, C组血管壁的tPAmRNA的表达量明显高于A组和B组(P<0. 01), 在术后 7d到达高峰, 并持续到 28d。扫描电镜观察显示, C组的血管壁只见少量血小板附着, 未见血栓形成; 而A组和B组的血管壁可见大量血小板黏附聚集, 且有血栓形成。免疫组化染色显示, C组的血管壁血小板源性生长因子(PDGF)阳性细胞的百分率明显低于A组和B组(P<0. 01)。A组和B组之间相比较, 以上数据差异无显著性(P> Objective: To investigate the effect of local transfection of tissue plasminogen activator (tPA) gene on intimal hyperplasia after injury of right external iliac artery in rabbits and to explore the possible mechanism. Methods: Microsurgery was used to establish the right external iliac artery injury model in rabbits. 105 New Zealand white rabbits were randomly divided into three groups of 35. Group A was saline control group, group B was liposome-mediated pBudCE4.1 transfection group, and group C was liposome-mediated pBudCE4.1 / tPA transfection group. Various transfection fluids were injected into the damaged vessel wall with a microinjector. Each group was subdivided into 5 subgroups, 7 rabbits in each subgroup, according to the experimental endpoint (2,3,7,14,28 days after surgery). At the end of each experiment, blood vessels of the injured section were taken for pathological examination, electron microscope observation, RT PCR and immunohistochemical staining. Results: Compared with group A and group B, the thickness of intima, the area of ​​intima and the stenosis of blood vessel in group C were significantly decreased at all time points after operation (P <0.01). At 28 days after operation, the stenosis rate of the lumen in group C was reduced by 51.5% and 54.2% respectively compared with that in group A and group B. At each time point, the expression of tPA mRNA in vessel wall of group C was significantly higher than that of group A and B (P <0.01), and reached the peak at 7 days after operation and lasted for 28 days. Scanning electron microscopy showed that only a small amount of platelet adhesion was seen in the vascular wall of group C, no thrombosis was seen. In the group A and group B, a large number of platelet adhesion and aggregation were observed in the blood vessel wall, and thrombosis occurred. Immunohistochemical staining showed that the percentage of platelet-derived growth factor (PDGF) positive cells in group C was significantly lower than that in group A and B (P <0.01). There was no significant difference in the above data between group A and group B (P>
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