金属蛋白涂层支架用于小型猪冠状动脉质粒介导下转基因研究(摘要)
作者：钱杰 高润霖 史瑞文 宋来凤 祈哲 李永利 魏英杰 孟亮 袁卫民 司文学 汤健
单位：北京市，中国医学科学院 中国协和医科大学 心血管病研究所；阜外心血管病医院 冠心病研究室(100037)
关键词：
中国循环杂志99zk47 目的：评价在质粒介导下，金属蛋白涂层支架向血管内局部转基因的可行性、效率和选择性。
方法：金属支架由316L不锈钢丝编织而成，载体为PcDNA2质粒，并携带有LacZ标记基因，该基因编码核特异性β-半乳糖苷酶。首先将蛋白涂层支架分别固定在3.0 mm或3.5 mm经皮冠状动脉腔内成形术球囊上并在浓度为8 μg/μl的基因原液中浸泡3分钟，吹干10分钟后通过8F大腔引导导管将支架送入小型猪冠状动脉前降支中段(n=3),另外把没有浸泡过基因的支架也送入小型猪冠状动脉前降支中段(n=3)。在支架置入后7天处死动物。β-半乳糖苷酶表达由X-Gal染色评估。
, http://www.100md.com
结果：所有转基因动物均有基因表达。转基因表达出现在内膜、中层和外膜。中层平滑肌细胞转染率为3.0%。远处器官和对照组冠状动脉均未显示核特异性β-半乳糖苷酶表达。
结论：蛋白涂层支架在质粒介导下向血管内转基因有效、可行，因此，它有可能成为冠状动脉腔内成形术后再狭窄基因治疗的有效转基因系统。
Plasmid-Mediated Intracoronary Local Gene Transfer Using
Protein-Coated Metallic Stent in Mini-Swine (Abstract)
Division of Coronary Heart Disease, Cardiovascular Institute and Fu Wai Hospital, CAMS and PUMC, Beijing (100037)
, 百拇医药
Qian Jie, Gao Runlin, Shi Ruiwen, et al.
Objective: To assess the feasibility, efficiency and selectivity of plasmid-mediated gene transfer to local arterial wall by protein-coated metallic stent.
Methods: Metallic stent was made by 316L stainless wire. The coating for metallic stent was made by immersing in gelatin solution containing crosslinker. PcDNA2 plasmid carrying LacZ reporter gene for nuclear-specific β-galactosidase was used. The coated stent was mounted on a 3.5 mm or 3.0 mm PTCA balloon and immersed into the gene stock (8 μg/μl) for 3 min, air-dried for 10 minutes and then implanted into the left anterior descending branch of coronary artery of mini-swine (n=3) via a 8F larger lumen guiding catheter. Coated stent without gene was implanted into the mini-swine of the some species as control (n=3). All the animals were sacrificed 7 days after stent implantation. β-galactosidase expression was assessed by X-gal staining.
, 百拇医药
Results: The expression of transgene was detected in local intima, media and adventitia of all the transgenic animals. The transfection efficiency of medial smooth muscle cells was 3.0%. No X-gal stained samples was detected in remote organs of the transgenic animals and neither in coronary arteries of the controls.
Conclusion: Plasmid-mediated arterial gene transfer to endothelial, smooth muscle cells and adventitia is feasible and efficacious with a protein-coated metallic stent. The coated stent may be a good carrier for plasmid-metdiated gene to be transferred and thus a potential tool to be used in transgenic prevention of restenosis following PTCA., 百拇医药
单位：北京市，中国医学科学院 中国协和医科大学 心血管病研究所；阜外心血管病医院 冠心病研究室(100037)
关键词：
中国循环杂志99zk47 目的：评价在质粒介导下，金属蛋白涂层支架向血管内局部转基因的可行性、效率和选择性。
方法：金属支架由316L不锈钢丝编织而成，载体为PcDNA2质粒，并携带有LacZ标记基因，该基因编码核特异性β-半乳糖苷酶。首先将蛋白涂层支架分别固定在3.0 mm或3.5 mm经皮冠状动脉腔内成形术球囊上并在浓度为8 μg/μl的基因原液中浸泡3分钟，吹干10分钟后通过8F大腔引导导管将支架送入小型猪冠状动脉前降支中段(n=3),另外把没有浸泡过基因的支架也送入小型猪冠状动脉前降支中段(n=3)。在支架置入后7天处死动物。β-半乳糖苷酶表达由X-Gal染色评估。
, http://www.100md.com
结果：所有转基因动物均有基因表达。转基因表达出现在内膜、中层和外膜。中层平滑肌细胞转染率为3.0%。远处器官和对照组冠状动脉均未显示核特异性β-半乳糖苷酶表达。
结论：蛋白涂层支架在质粒介导下向血管内转基因有效、可行，因此，它有可能成为冠状动脉腔内成形术后再狭窄基因治疗的有效转基因系统。
Plasmid-Mediated Intracoronary Local Gene Transfer Using
Protein-Coated Metallic Stent in Mini-Swine (Abstract)
Division of Coronary Heart Disease, Cardiovascular Institute and Fu Wai Hospital, CAMS and PUMC, Beijing (100037)
, 百拇医药
Qian Jie, Gao Runlin, Shi Ruiwen, et al.
Objective: To assess the feasibility, efficiency and selectivity of plasmid-mediated gene transfer to local arterial wall by protein-coated metallic stent.
Methods: Metallic stent was made by 316L stainless wire. The coating for metallic stent was made by immersing in gelatin solution containing crosslinker. PcDNA2 plasmid carrying LacZ reporter gene for nuclear-specific β-galactosidase was used. The coated stent was mounted on a 3.5 mm or 3.0 mm PTCA balloon and immersed into the gene stock (8 μg/μl) for 3 min, air-dried for 10 minutes and then implanted into the left anterior descending branch of coronary artery of mini-swine (n=3) via a 8F larger lumen guiding catheter. Coated stent without gene was implanted into the mini-swine of the some species as control (n=3). All the animals were sacrificed 7 days after stent implantation. β-galactosidase expression was assessed by X-gal staining.
, 百拇医药
Results: The expression of transgene was detected in local intima, media and adventitia of all the transgenic animals. The transfection efficiency of medial smooth muscle cells was 3.0%. No X-gal stained samples was detected in remote organs of the transgenic animals and neither in coronary arteries of the controls.
Conclusion: Plasmid-mediated arterial gene transfer to endothelial, smooth muscle cells and adventitia is feasible and efficacious with a protein-coated metallic stent. The coated stent may be a good carrier for plasmid-metdiated gene to be transferred and thus a potential tool to be used in transgenic prevention of restenosis following PTCA., 百拇医药