Department of Cardiovascular Surgery, University Hospital of Berne, Berne, Switzerland.
PLoS One. 2010 Dec 30;5(12):e14467. doi: 10.1371/journal.pone.0014467.
Gene therapy may represent a promising alternative strategy for cardiac muscle regeneration. In vivo electroporation, a physical method of gene transfer, has recently evolved as an efficient method for gene transfer. In the current study, we investigated the efficiency and safety of a protocol involving in vivo electroporation for gene transfer to the beating heart. Adult male rats were anesthetised and the heart exposed through a left thoracotomy. Naked plasmid DNA was injected retrograde into the transiently occluded coronary sinus before the electric pulses were applied. Animals were sacrificed at specific time points and gene expression was detected. Results were compared to the group of animals where no electric pulses were applied. No post-procedure arrhythmia was observed. Left ventricular function was temporarily altered only in the group were high pulses were applied; CK-MB (Creatine kinase) and TNT (Troponin T) were also altered only in this group. Histology showed no signs of toxicity. Gene expression was highest at day one. Our results provide evidence that in vivo electroporation with an optimized protocol is a safe and effective tool for nonviral gene delivery to the beating heart. This method may be promising for clinical settings especially for perioperative gene delivery.
基因治疗可能代表了心肌再生的一种有前途的替代策略。体内电穿孔是一种基因转移的物理方法，最近已发展成为一种高效的基因转移方法。在本研究中，我们研究了一种涉及体内电穿孔的方案，该方案将基因转移到跳动的心脏，以评估其效率和安全性。成年雄性大鼠被麻醉，通过左开胸术暴露心脏。在施加电脉冲之前，将裸露的质粒 DNA 逆行注射到短暂闭塞的冠状窦中。在特定时间点处死动物，并检测基因表达。将结果与未施加电脉冲的动物组进行比较。没有观察到术后心律失常。仅在施加高脉冲的组中观察到左心室功能暂时改变；仅在该组中 CK-MB（肌酸激酶）和 TNT（肌钙蛋白 T）也发生改变。组织学检查没有显示出毒性迹象。基因表达在第 1 天最高。我们的结果提供了证据，表明使用优化方案的体内电穿孔是将非病毒基因递送到跳动心脏的一种安全有效的工具。这种方法对于临床环境，特别是围手术期基因传递可能很有前途。
