Chikada M, Jones M
Department of Cardiovascular Surgery, National Children's Hospital, Tokyo, Japan.
Jpn J Thorac Cardiovasc Surg. 1999 May;47(5):204-9. doi: 10.1007/BF03217995.
Gene therapy is a therapeutic strategy in treating cardiovascular disease. Vein graft failure, the major limitation on coronary artery bypass surgery, may be amenable to gene approaches. Some studies describe gene therapies using functioning genes to prevent vein graft stenosis. Gene transfer efficiency remains a major issue. In this rabbit vein graft model, we studied gene delivery using a replication-deficit recombinant adenovirus to improve gene transfer efficiency into vein grafts. The adenovirus vector that contains the E.coli lacZ gene encoding beta gal was used because this vector is widely used and thought to be effective. Gene transfer was detected by X-gal staining. We hypothesized that dimethylsulfoxide and hyaluronidase, both drug delivery enhancers, would improve efficiency and that, in transfer to adventitia, direct injection would be more effective than dwelling. We studied 3 gene delivery methods to intima and media (controls, using dimethylsulfoxide and using hyaluronidase before transfection) and 2 delivery methods to adventitia (direct injection and dwelling). We used 6 rabbits per delivery method. X-gal stained positive cell rates were counted using light microscopy. Our findings indicate that (1) dimethylsulfoxide increased the efficiency of transfection to media and intima, (2) hyaluronidase increased intimal transfection efficiency, (3) direct injection to adventitia was more effective than dwelling. These findings suggest that in vein grafting, our methods are feasible for improving gene transfer efficiency.
基因治疗是治疗心血管疾病的一种治疗策略。静脉移植物失败是冠状动脉搭桥手术的主要限制因素，可能适合采用基因治疗方法。一些研究描述了使用功能基因的基因疗法来预防静脉移植物狭窄。基因转移效率仍然是一个主要问题。在这个兔静脉移植物模型中，我们研究了使用复制缺陷型重组腺病毒进行基因递送，以提高基因向静脉移植物中的转移效率。使用了含有编码β-半乳糖苷酶的大肠杆菌lacZ基因的腺病毒载体，因为该载体被广泛使用且被认为是有效的。通过X-gal染色检测基因转移。我们假设，作为两种药物递送增强剂的二甲亚砜和透明质酸酶会提高效率，并且在向 adventitia 转移时，直接注射比留置更有效。我们研究了3种向内膜和中膜递送基因的方法（对照组、使用二甲亚砜以及在转染前使用透明质酸酶）和2种向 adventitia 递送基因的方法（直接注射和留置）。每种递送方法使用6只兔子。使用光学显微镜计算X-gal染色阳性细胞率。我们的研究结果表明：（1）二甲亚砜提高了向中膜和内膜的转染效率；（2）透明质酸酶提高了内膜转染效率；（3）向 adventitia 直接注射比留置更有效。这些发现表明，在静脉移植中，我们的方法对于提高基因转移效率是可行的。
