Intravascular local gene transfer mediated by protein-coated metallic stent  

金属支架蛋白涂层所介导的血管内局部转基因(英文)

作　　者：袁晋青[1] 高润霖[1] 史瑞文[1] 宋来风[1] 汤健[1] 李永力[1] 唐承君[1] 孟亮[1] 袁卫民[1] 陈在嘉[1] 

机构地区：[1]中国医学科学院中国协和医科大学阜外心血管病医院冠心病研究室,北京100037

出　　处：《Chinese Medical Journal》2001年第10期35-37,105,共4页中华医学杂志（英文版）

基　　金：ThisstudywassupportedbyagrantfromNationalResearchKeyProjectoftheNinth FiveYearPlan (No 96 90 6 0 2 0 7)

摘　　要：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 the Lac Z reporter gene for nuclearspecific β-galactosidase (Ad-βgal) was used in this study. The coating for metallic stent was made by immersing it in a gelatin solution containing crosslinker. The coated stents were mounted on a 4.0 or 3.0 mm percutaneous transluminal coronary angioplasty (PTCA) balloon and submersed into a high-titer Ad-βgal viral stock (2 × 1010 pfu/ml) for 3 min, and then implanted into the carotid artedes in 4 mini-swines and into the left anterior descending branch of the coronary artery in 2 mini-swines via 8F large lumen guiding catheters. The animals were sacrificed 7 (n=4), 14 (n = 1) and 21 (n = 1 ) days after implantation, respectively. The β-galactosidase expression was assessed by X-gal staining.Results The results showed that the expression of transgene was detected in all animal. In 1 of carotid artery with an intact intima, the β-gal expression was limited to endothelial cells. In vessels with denuded endothelium, gene expression was found in the sub-intima, media and adventitia. The transfection efficiency of medial smooth muscle cells was 38.6%. In 2 animals sacrificed 7 days after transfection, a microscopic examination of X-gal-stained samples did not show 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 by protein-coated metallic stent is feasible. The transfection efficiency is higher. The coated stent may act as a good carrier of adenovirus-mediated gene transfer and have a potential to prevent restenosis following PTCA.目的　评估在腺病毒介导下 ,由交联蛋白涂层金属支架向局部动脉壁转基因的可行性和效率。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。首先将交联蛋白涂层支架分别固定在 4 0和 3 0mmPTCA球囊上并在高效价病毒原液中 (2× 10 10 pfu/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 ,球囊以 10 - 12个大气压扩张二次 ,每次 1分钟。在支架植入后 7(n =4 )、14(n =1)和 2 1(n =2 )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　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。该涂层支架可做为腺病毒介导转基因的良好运载体 ,并有可能预防PTCA术后再狭窄。

关 键 词：gene transfer  ·  stent   ·  restenosis  ·   adenovirus 

分 类 号：R540.5[医药卫生—心血管疾病]