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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?
Objective To observe the characteristics of the carotid artery transfected with adenoviral vector in rats, namely, the efficiency of gene transfection and the time course of exogenous gene expression, and to explore the effects of local vascular transgene on various organs of the body. Methods The rats were subjected to carotid artery anastomosis with a soluble stent. The Adv5 CMV plasmid (control group) or Adv5 CMV / LacZ plasmid (treatment group) was transfused into the carotid artery of rats. The time of transfection (stent implantation to recovery of blood flow) minute. Carotid arteries were obtained at 2, 7, 14, 28, 60 and 90 days after operation. The ascending aorta, heart, brain, liver, lung, Glycosidase (expression product of LacZ gene) activity assay and histochemical staining. Results (1) β-galactosidase activity assay: All the anastomosis carotid arteries in the control group had no β-galactosidase activity at 2, 7, 14, 28, 60 and 90 days after operation; Β - galactosidase gene expression, 7 to 14 days for the peak of gene expression, 28 days after the gene expression decreased significantly, but low levels of gene expression can last for 3 months. The ascending aorta, heart, brain, liver, lung, spleen and kidney of the treated group had no β - galactosidase activity at 7 and 14 days after operation. (2) The results of histochemical staining of β - galactosidase: The blue dye was observed in each anastomosis carotid artery in the treatment group at 2, 7, 14, 2, 8, 60 and 90 days after operation, Carotid transverse cross-section of the whole week are blue dye, the inner layer of the blood vessel wall, the middle part of the majority of cells are dyed blue. The ascending aorta, heart, brain, liver, lung, spleen and kidney of the treated group were not blue-stained at 7 and 14 days after operation.