腺病毒载体转染局部血管的安全性研究

Safety of direct gene transfer into rat carotid arteries mediated by adenoviral vector

目的 观察腺病毒载体转染局部血管，外源性基因的表达能否局限于转染部位，而不出现于其他部位，亦即观察腺病毒载体转染局部血管的安全性。 方法 用可溶性支架将Ａｄｖ５ＣＭＶ或Ａｄｖ５ＣＭＶ／ＬａｃＺ质粒导入１６ 只大鼠颈动脉吻合口。术后７ 天，取对照组和治疗组全部已经行吻合的颈动脉以及治疗组的升主动脉、心、脑、肝、肺、脾、肾，行β半乳糖苷酶活性测定和组织化学染色。 结果 治疗组颈动脉β半乳糖苷酶表达量为１０－１－３ ×１０ｉｕ／ｇ 组织，每条颈动脉横切面的全周均可见蓝染。对照组颈动脉及治疗组升主动脉、心、脑、肝、肺、脾、肾均未测到β半乳糖苷酶活性，组化染色未见蓝染细胞。 结论 用腺病毒载体转染局部血管，虽然转染血管局部有外源性基因的高水平表达，但该表达仅局限于转染部位，不出现于其他脏器，亦即具有良好的安全性。

Objective To evaluate the safety of direct gene transfer into rat carotid arteries mediated by adenoviral vector. Methods After being soaked in high concentration of glucose solution containing Adv5 CMV (control group) or Adv5 CMV/LacZ(treatment group) for 30 minutes, soluble stents were inserted into the lumina of rat carotid arteries and end to end anastomoses of cut rat carotid were performed with standard microvascular surgical technique. On the 7th day after surgery, anastomotic carotid arteries of the two groups were obtained, and samples of ascending aortas, hearts, brains, livers, lungs, spleens and kidneys of the treatment group were taken for assessing β galactosidase activity and histochemical staining. Results β galactosidase activity was not detected in the carotid arteries in the control group and in those organs not directly exposed to adenoviral vector in the treatment group. The amount of β galactosidase expression in the carotid arteries in the treatment group was 10.1×10 -3 IU/g tissue. Microscopic examination of sections from vessels in the control group and from the aorta, heart, brain, liver, lung, spleen or kidney in the treatment group revealed no X gal staining. Microscopic examination of the carotid arteries in the treatment group revealed blue staining in all anastomotic arteries and in all layers of the arterial wall, including the intima, media, and adventitia. Conclusions A recombinant gene could be safely delivered to a local site of the vessel by means of infection with adenoviral vector.