Treatment of acute limb ischemia by intramuscular injection of vascular endothelial growth factor gene.
Treatment of acute limb ischemia by intramuscular injection of vascular endothelial growth factor gene.
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发表时间：
1997-11
期刊：
影响因子：
37.8
通讯作者：
Y. Tsurumi;M. Kearney;Dongfen Chen;Marcy Silver;S. Takeshita;Jihong Yang;J. Symes;J. Isner
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文献类型：
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作者：
Y. Tsurumi;M. Kearney;Dongfen Chen;Marcy Silver;S. Takeshita;Jihong Yang;J. Symes;J. Isner
背景缺血性骨骼肌已被证明有利于摄取和表达以裸质粒DNA形式转移的基因。因此，急性缺血的骨骼肌可能代表裸DNA IM基因治疗的潜在靶点。因此，我们研究了肌内注射编码分泌性血管生成生长因子、血管内皮生长因子（VEGF）的质粒DNA对急性后肢缺血动物模型中侧支血管发育的影响。方法与结果新西兰白兔远端髂外动脉结扎后，直接向缺血的大腿肌肉注射500μg phVEGF165。转染后 30 天，与对照组相比，VEGF 转染动物具有更多血管造影可识别的侧支血管（血管造影评分=0.72+/-0.06 与 0.48+/-0.10；P<.01）以及组织学评估的毛细血管（248+/-37 与 180+/-32/mm2，P<.01）。通过小腿收缩压比（0.80+/-0.09 与 0.56+/-0.10，P<.01）和用多普勒导丝测量的缺血肢体血流（静息血流=22+/-5 与 14+/-4；P<.01；充血血流=59+/-17 与 39+/-12 mL/min；充血血流=59+/-17 与 39+/-12 mL/min； P<.05)。基因转移后长达 14 天，人 VEGF mRNA 在转染的缺血肌肉中表达。根据报告质粒表达，缺血肌肉中的转染效率比非缺血对照肌肉中高六倍。结论 这些结果表明采用直接 IM 转移裸露 VEGF 质粒 DNA 来优化急性肢体缺血治疗的可行性。
BACKGROUND Ischemic skeletal muscle has been shown to be advantageous for taking up and expressing genes transferred in the form of naked plasmid DNA. Therefore, acutely ischemic skeletal muscle may represent a potential target for IM gene therapy with naked DNA. Accordingly, we investigated the impact of IM injection of plasmid DNA encoding the secreted angiogenic growth factor, vascular endothelial growth factor (VEGF), on collateral vessel development in an animal model of acute hindlimb ischemia. METHODS AND RESULTS After ligation of distal external iliac artery in New Zealand White rabbits, we directly injected 500 microg of phVEGF165 into the ischemic thigh muscles. At 30 days posttransfection, VEGF-transfected animals had more angiographically recognizable collateral vessels (angiographic score=0.72+/-0.06 versus 0.48+/-0.10; P<.01) as well as histologically assessed capillaries (248+/-37 versus 180+/-32/mm2, P<.01) compared to controls. Hemodynamic deficit was less severe in VEGF-transfected animals by calf systolic blood pressure ratio (0.80+/-0.09 versus 0.56+/-0.10, P<.01) and by flow to the ischemic limb measured with Doppler guidewire (resting flow=22+/-5 versus 14+/-4; P<.01; hyperemic flow=59+/-17 versus 39+/-12 mL/min; P<.05). Human VEGF mRNA was expressed in the transfected ischemic muscles as long as 14 days after gene transfer. Based on reporter plasmid expression, transfection efficiency was sixfold higher in ischemic muscles than in nonischemic control muscles. CONCLUSIONS These results suggest the feasibility of employing direct IM transfer of naked VEGF plasmid DNA to optimize treatment of acute limb ischemia.