Optimization of adenovirus-mediated endothelial nitric oxide synthase delivery in rat hindlimb ischemia.
Optimization of adenovirus-mediated endothelial nitric oxide synthase delivery in rat hindlimb ischemia.
复制标题
大鼠后肢缺血中腺病毒介导的内皮一氧化氮合酶递送的优化。
DOI：
10.1038/sj.gt.3302563
复制
发表时间：
2005
期刊：
影响因子：
5.1
通讯作者：
Messina,LM
中科院分区：
文献类型：
--
作者：
Yan,J;Tang,GL;Wang,R;Messina,LM
腺病毒介导的内皮型一氧化氮合酶（eNOS）过表达诱导侧支动脉发育，并在诱导实验性急性后肢缺血后显著增加血流量。然而，对于这种或任何其他形式的肢体缺血基因治疗的最佳基因递送技术仍然是未知的。本研究的目的是确定两种最常用的技术，动脉内和肌内注射，对血流恢复，侧支动脉发育和肌肉质量保存的影响。我们比较了血管隔离下的动脉内注射，短暂血管闭塞下的动脉内注射，以及诱导急性后肢缺血后肌肉内注射磷酸盐缓冲盐水（PBS）或编码eNOS（AdeNOS）或LacZ（AdlacZ）基因的腺病毒。与肌肉注射相比，动脉注射AdeNOS可增加eNOS活性（22.30 vs. 10.56，P< 0.01）、血流量（0.90±0.02 vs. 0.69±0.07，P< 0.001）和侧支动脉发育（17.56484 vs. 13.74259，P< 0.05）。短暂血管闭塞下的动脉内注射导致更好地保存肌肉质量，肌肉结构和临床缺血指数，但导致更大的转基因表达在远端器官和对侧肢体肌肉。在短暂的血管闭塞下动脉内注射AdeNOS是逆转大鼠严重后肢缺血的最佳技术。这是第一次系统地比较不同的传递技术用于实验性后肢缺血的基因治疗。
Adenovirus-mediated overexpression of endothelial nitric oxide synthase (eNOS) induces collateral artery development and substantially increases blood flow after induction of experimental acute hindlimb ischemia. However, the optimal technique of gene delivery for this or any other form of gene therapy in limb ischemia is still unknown. The purpose of this study was to determine the effect of the two most commonly used techniques, intra-arterial and intramuscular injection, on blood flow recovery, collateral artery development, and preservation of muscle mass. We compared intra-arterial injection under vascular isolation, intra-arterial injection under transient vascular occlusion, and intramuscular injection of phosphate buffered saline (PBS) or adenovirus encoding either the eNOS (AdeNOS) or LacZ (AdlacZ) gene after induction of acute hindlimb ischemia. Delivery of AdeNOS by both intra-arterial injection techniques increased eNOS activity (22.30 versus 10.56, P< 0.01), blood flow (0.90±0.02 versus 0.69±0.07, P< 0.001) and collateral artery development (17.56484 versus 13.74259, P< 0.05) more than by intramuscular delivery. Intra-arterial injection under transient vascular occlusion led to better preservation of muscle mass, muscle architecture, and clinical ischemic index, but led to greater transgene expression in distant organs and contralateral limb muscles. Intra-arterial injection of AdeNOS under transient vascular occlusion is the optimal technique to reverse severe hindlimb ischemia in the rat. This is the first systematic comparison of different delivery techniques used in gene therapy of experimental hindlimb ischemia.