生物蛋白胶外支架提高血管外膜腺病毒转染效率的实验研究

Fibrin Glue as External Support Enhanced Adventitial Adenovirus Transfection

目的:将携带报告基因的腺病毒载体与生物蛋白胶混合,喷洒在移植静脉外膜,以观察生物蛋白胶是否能提高血管外膜基因转染效率。方法:健康纯种新西兰大耳白兔,随机分为两组单纯腺病毒对照组和生物蛋白胶腺病毒组,每组8只兔。建立兔颈外静脉颈总动脉旁路移植术模型。对照组移植静脉外周单独给予携带β-半乳糖苷酶报告基因的腺病毒载体,生物蛋白胶腺病毒组则给予生物蛋白胶腺病毒载体混合物。分别于术后第7天和第14天取出移植静脉,评价转染效率。结果:术后第7天,对照组腺病毒转染效率(2.1%±0.5%),而生物蛋白胶腺病毒组表现为高转染效率(13.2%±4.6%),两者比较有极显著差异;术后第14天,对照组偶见阳性转染细胞,而在生物蛋白胶腺病毒组虽然阳性转染细胞较术后第7天为少,但仍保持高转染率(6.3%±3.8%)。结论:生物蛋白胶作为一种新的药物缓释系统,能明显提高移植静脉外膜基因转染效率。

Objective： Intraluminal gene therapy is limited by inadequate exposure time due to run-off associated with blood flow and prolonged ischemia time which would worsen intimal injury. Peri-adventitial gene delivery is a promising alternative therapy to reduce intimal hyperplasia. But it is limited by low and transient levels of gene transfection. To get more effective gene transfection through peri-adventitial, a study was undertaken to investigate whether mixing adenovirus with fibrin glue may increase the level and prolong the time period of gene expression. Methods： Right jugular vein to common carotid artery interposition grafting was performed in sixteen male New Zealand white rabbits （2.5-3.0 kg） and the animals were divided into two groups： Ad-GAL group （n = 8）; fibrin glue/Ad-GAL group （n = 8）. Commercially available fibrin glue and adenovirus expressing the gene for β-galactosidase （Ad-GAL） was applied separately or in mixing around vein grafts. At 7th day and 14th day after implantation, the grafts were harvested to evaluate transfection rate. Results： Compared with weak staining in 2. 1 % ± 0.5%in Ad-GAL alone grafts, a high level of β-Galactosidase staining was evident in 13.2%±4.6% in fibrin glue/Ad-GAL grafts at 7th day（P〈0. 001）. At 14th day, almost no staining （0G） was detected in Ad-GAL alone grafts. However, there was still a relative high level staining （6.3%±3.8%） in fibrin glue/Ad-GAL grafts （P〈0. 001） vs. Ad-GAL alone group（P = 0.01 ）. Conclusion： A novel method of adventitial gene delivery using fibrin glue as external support is proposed. Fibrin glue may be an ideal candidate for controlled release delivery which would facilitate adventitial gene transfer.