Veit K, Boissel J P, Buerke M, Grosser T, Meyer J, Darius H
Department of Medicine II, Johannes Gutenberg-University, Mainz, Germany.
Cardiovasc Res. 1999 Aug 15;43(3):808-22. doi: 10.1016/s0008-6363(99)00172-8.
The efficient introduction of regulatory genes into vascular smooth muscle cells (SMCs) is one of the most promising options for gene therapy of cardiovascular diseases. Cationic liposome-mediated gene transfer may become a favorable transfection technique with regard to patient's safety for in vivo administration. However, this method until now has its limitation in a low transfection efficiency. Therefore, the present study was designed to improve cationic liposome-mediated transfection of rabbit vascular SMCs in vitro and in vivo, in order to enhance transfection efficiency and present an optimized system which may offer a potential therapeutic benefit for in vivo application.
Optimized lipofection of rabbit SMCs with the mammalian expression vector pE-N1 and the reporter gene green fluorescent protein resulted in a mean transfection efficiency of about 50%. The unique transfection of rabbit SMCs in vitro and in vivo with the inducible isoform of human nitric oxide synthase (NOSII), using the same vector, resulted in a successful transient transcription and translation of a functionally active human NOSII in rabbit SMC, persisting 5-6 days. We could further demonstrate that the transfection procedure and the transgene product did neither induce necrosis nor apoptosis under the conditions chosen and did not result in the induction of endogenous NOSII of transfected SMCs.
CONCLUSION(S): These findings indicate potential therapeutic relevance for this nonviral gene transfer system for in vivo gene therapy for cardiovascular diseases.
将调控基因有效导入血管平滑肌细胞（SMC）是心血管疾病基因治疗最有前景的方法之一。阳离子脂质体介导的基因转移就体内给药患者安全性而言可能成为一种理想的转染技术。然而，目前该方法存在转染效率低的局限性。因此，本研究旨在提高阳离子脂质体介导的兔血管SMC体外和体内转染效率，以增强转染效果并提供一种优化系统，为体内应用提供潜在治疗益处。
用哺乳动物表达载体pE-N1和报告基因绿色荧光蛋白对兔SMC进行优化脂质体转染，平均转染效率约为50%。使用相同载体在体外和体内对兔SMC进行独特转染，转染人诱导型一氧化氮合酶（NOSII），结果在兔SMC中成功实现了功能性活性人NOSII的瞬时转录和翻译，持续5至6天。我们还能进一步证明，在所选择的条件下，转染过程和转基因产物既不诱导坏死也不诱导凋亡，也不会导致转染SMC的内源性NOSII的诱导。
这些发现表明这种非病毒基因转移系统在心血管疾病体内基因治疗中具有潜在的治疗意义。
