Gene transfer and expression in rat anastomotic artery in vivo using adenoviral vector  

腺病毒载体转染在体血管的特性(英文)

作　　者：黄志雄[1] 郭加强[1] 郭少先[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心血管病研究所,北京100037

出　　处：《Chinese Medical Journal》2001年第6期90-93,110-111,共6页中华医学杂志（英文版）

摘　　要：Objective　To observe the efficiency and time course of gene expression and the safety of adenoviral vector mediated gene transfer in vivo.Methods　After soaking soluble stents in a high concentration of glucose solution containing Adv5-CMV (cytomegalovirus) (control group) or Adv5-CMV/LacZ (treatment group) for 30 minutes, the stents were inserted into the lumina of cut rat carotid arteries and end-to-end anastomoses of the cut carotid were performed with standard microvascular surgical techniques. On days 2, 7, 14, 28, 60 and 90 after gene transfer, anastomotic arteries of the two groups were observed. On days 7 and 14, the ascending aortas, hearts, brains, livers, lungs, spleens and kidneys of the treatment group were observed. All samples were analyzed for the presence of β-galactosidase activity and histochemical staining.Results　β-galactosidase activity was not detected in the carotid arteries of the control group and organs not directly exposed to adenoviral vector of the treatment group. The amount of β-galactosidase activity (×10-3?U/g tissue) in the treatment group on the 2nd, 7th, 14th, 28th, 60th and 90th day after gene transfer was 3.87, 11.38, 9.8, 6.43, 3.18 and 2.43, respectively. Microscopic examination of sections from vessels of the control group and from the aortas, hearts, brains, livers, lungs, spleens or kidneys of the treatment group revealed no X-gal staining. Microscopic examination of carotid arteries of the treatment group revealed blue-staining in all anastomotic arteries and in all layers of the arterial wall observed on days 7 and 14 after gene transfer.Conclusion　Adenoviral vector can effectively infect blood vessels in vivo. After adenoviral vector mediated direct gene transfer into anastomotic rat carotid arteries, recombinant gene expression began on day 2, peaked between days 7 and 14, prominently declined after day 28, and persisted at low levels more than three months. A recombinant gene could be delivered to a specific site by direct gene transfer in vivo by adenov目的　观察腺病毒载体转染大鼠颈动脉的特性即基因转染的有效性和外源性基因表达的时间过程以及探讨局部血管转基因对全身各脏器的影响。方法　用可溶性支架行大鼠颈动脉吻合 ,使Adv5 CMV质粒 (对照组 )或Adv5 CMV/LacZ质粒 (治疗组 )转染大鼠颈动脉 ,转染时间 (置入支架至恢复血流 )为 30分钟。术后 2、7、14、2 8、60及 90天取吻合的颈动脉 ,术后 7、14天取治疗组升主动脉、心、脑、肝、肺、脾、肾 ,行β 半乳糖苷酶 (LacZ基因的表达产物 )活性测定和组织化学染色。结果　 ( 1) β 半乳糖苷酶活性测定 :对照组术后 2、7、14、2 8、60、90天所有吻合颈动脉均无β 半乳糖苷酶活性 ;治疗组术后第 2天即有β -半乳糖苷酶基因的表达 ,7∽ 14天为基因表达的高峰期 ,2 8天后基因表达量明显减少 ,但低水平的基因表达可持续 3个月之久。治疗组术后 7、14天所取升主动脉、心、脑、肝、肺、脾及肾组织均无 β -半乳糖苷酶活性。( 2 ) β -半乳糖苷酶组织化学染色结果 :治疗组术后 2、7、14、2 8、60、90天每条吻合颈动脉均可见蓝染 ,其中术后 7、14天每条颈动脉的横切面的全周均有蓝染 ,血管壁内层、中层的大部分细胞被染成蓝色。治疗组术后 7、14天所取升主动脉、心、脑、肝、肺、脾及肾组织均无蓝染 ,对照组所?

关 键 词：genetic vectors  ·   gene transfer  ·   braces  ·   gene expression 

分 类 号：R373[医药卫生—病原生物学]