金属支架蛋白涂层介导的血管内局部转基因  

Intravascular Local Gene Transfer Mediated by Protein coated Metallic Stent

作　　者：袁晋青[1,2,3] 高润霖[1,2,3] 史瑞文[1,2,3] 宋来凤 李永利[1,2,3] 孟亮 袁卫民[1,2,3] 司文学 汤健[1,2,3] 

机构地区：[1]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 [2]中国医学科学院生物医学工程研究所 [3]北京医科大学心血管研究所

出　　处：《中国循环杂志》1997年第5期330-333,共4页Chinese Circulation Journal

摘　　要：目的：评估在腺病毒介导下，由蛋白涂层金属支架向局部动脉壁转基因的可行性、效率和选择性。方法：金属支架由３１６Ｌ不锈钢丝编织而成，载体为复制缺陷的重组腺病毒，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４．０和３．０ｍｍ经皮冠状动脉腔内成形术球囊上并在高效价病毒原液中（２×１０１０ｐｆｕ／ｍｌ）浸泡３分钟，然后通过８Ｆ大腔引导导管分别将支架植入小型猪颈动脉（ｎ＝４）和冠状动脉前降支中段（ｎ＝２）。在支架植入后７（ｎ＝４），１４（ｎ＝１）和２１（ｎ＝１）天分别将动物处死。β－半乳糖苷酶表达由Ｘ－Ｇａｌ染色评估。在转染后７天行组织化学染色检测远处器官转基因表达。结果：所有动物均发现转基因表达。在１例有完整内膜的颈动脉，β－半乳糖苷酶表达局限于内皮细胞，而当内皮剥落后，转基因表达出现在内膜下、中层和外膜。中层平滑肌细胞转染率为３８．６％。远处器官和邻近支架的近远端血管及对照血管均未显示核特异性β－半乳糖苷酶的表达。结论：①在腺病毒介导下，由交联蛋白涂层金属支架向内皮，中层平滑肌细胞和外膜转基因有效可行且转染效率较高。②该涂层支架可做为腺病毒介导转基因的良好运载体，并有可能?Objective:To assess the feasibility,efficiency and selectivity of adenovirus mediated gene transfer to local arterial wall by protein coated metallic stent. Methods:A replication defective recombinant adenovirus carrying LacZ reporter gene for nuclear specific β galactosidase(Ad β gal) was used in this study.The coating for metallic stent was made by immersing it in gelatin solution containing crosslinker.The coated stents were mounted on 4 0 and 3 0mm PTCA balloon and submersed into high titer Ad β gal viral stock(2×10 10 pfu/ml) for 3 min,then implanted into carotid arteries in 4 mini swine and left anterior descending branch of coronary artery in 2 mini swine via 8F large lumen guiding catheters.The animals were sacrificed respectively 7( n =4),14( n =1)and 21( n =1)days after implantation.The β galactosidase expression was assessed by x gal staining. Results:The expression of transgene was detected in all animals.In 1 carotid artery with intact intima,the β gal expression was limited to endothelial cells.In vessels with denuded endothelium gene expression was found in sub intima,media and adventitia.The transfection efficiency of medial smooth muscle cells was 38 6%.In 2 animals sacrificed 7 days after coronary transfection,microscopic examination of X gal stained samples failed to detect evidence of transfection in remote organs and arterial segments adjacent to the treated arterial site. Conclusions:Adenovirus mediated arterial gene transfer to endothelial,smooth muscle cells and adventitia is feasible by protein coated metallic stent with a high transfection efficiency.The coated stent may act as a good carrier of adenovirus mediated gene transfer and have a potential to prevent the restenosis following PTCA.

关 键 词：转基因 腺病毒 金属支架 蛋白涂层 冠状动脉狭窄 

分 类 号：R543.305[医药卫生—心血管疾病] R654.3[医药卫生—内科学]