腺病毒载体转染大鼠颈动脉的实验研究

目的观察腺病毒载体转染大鼠颈动脉的有效性和外源性基因表达的时间过程。方法用ＡｄＶ５－ＣＭＶ质粒（对照组）或Ａｄｖ５－ＣＭＶ／ＬａｃＺ质粒（治疗组）转染大鼠颈动脉３０分钟。术后２、７、１４、２８、６０及９０天取被转染的颈动脉,行β－半乳糖苷酶活性测定和组织化学染色。结果对照组所有颈动脉均无β－半乳糖苷酶活性,治疗组术后２天颈动脉即有β－半乳糖苷酶的表达,７～１４天为表达高峰期,２８天后表达量明显减少,但可持续表达３个月之久。治疗组术后２、７、１４、２８、６０及９０天每条颈动脉均可见蓝染,其中术后７及１４天每条颈动脉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。对照组所有动脉均未见蓝染。结论腺病毒载体能高效转染大鼠在体颈动脉。转染后,外源性基因的高水平表达只能维持１个月左右。

Gene expression in vivo mediated by adenoviral vector after direct gene transfer into anastomotic rat carotid arteries

OBJECTIVE: To observe the efficiency and time course of gene expression in vivo mediated by adenoviral vector after direct gene transfer into anastomotic rat carotid arteries. 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 cut rat carotid arteries and end-to-end anastomoses were performed. On the 2nd, 7th, 14th, 28th, 60th and 90th day after surgery, the anastomotic carotid arteries of the two groups were explanted. All samples were analyzed for the presence of beta-galactosidase activity and histochemical staining. RESULTS: beta-galactosidase activity was not detected in the carotid arteries of the control group. Microscopic examination of sections from vessels of the control group revealed no X-gal staining. The treatment group revealed bluestaining in all anastomotic arteries and in all layers of the arterial wall explanted including the intima, media, and adventitia on the 7th, 14th day postoperatively. CONCLUSIONS: Adenoviral vector could make foreign effectively transfer into rat carotid arteries in vivo. Mediated by adenoviral vector after direct gene transfer into anastomotic rat carotid arteries, the high level of gene expression only persisted about one month.