Overexpression of endothelial nitric oxide synthase increases skeletal muscle blood flow and oxygenation in severe rat hind limb ischemia
Overexpression of endothelial nitric oxide synthase increases skeletal muscle blood flow and oxygenation in severe rat hind limb ischemia
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DOI：
10.1016/s0741-5214(03)00555-x
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发表时间：
2003-10-01
影响因子：
4.3
通讯作者：
Messina, LM
中科院分区：
文献类型：
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作者：
Brevetti, LS;Chang, DS;Messina, LM
Objective.虽然一氧化氮（NO）在血管生成中具有关键作用，但NO合酶过表达在严重缺血中的治疗潜力仍不明确。我们测试的假设，过度表达内皮型一氧化氮合酶（eNOS）将改善组织灌注在严重的后肢ischemia.Methods：严重的后肢缺血诱导122成年雄性SD大鼠。诱导后肢缺血后10天，进行血管分离和动脉内递送编码eNOS的腺病毒载体（AdeNOS）、对照腺病毒载体（AdE1）或磷酸盐缓冲盐水溶液（PBS）。结果：AdeNOS基因转染后，eNOS蛋白表达和酶活性增加，血管造影和毛细血管计数免疫组化显示eNOS蛋白表达和酶活性增加。基因转移后两周，eNOS转导的大鼠骨骼肌血流量比AdE1转导或PBS处理的大鼠高4倍，与非缺血性肌肉中运动诱导的最大血流量相似。eNOS过表达以滴度依赖性方式增加肌氧张力。这种增加持续1个月后转导，即使eNOS酶活性下降到正常水平。血管造影和毛细血管计数表明，eNOS过度表达增加侧支动脉的大小和数量，但没有显着增加毛细血管肌纤维ratio.Conclusions：eNOS过度表达在缺血大鼠后肢显着增加骨骼肌血流量，肌氧分压，侧支动脉（动脉生成）。此外，eNOS过表达没有导致高于对照水平的毛细血管生成。这些研究证明了eNOS过表达作为人类严重肢体缺血治疗的潜力。
Objective. Although nitric oxide (NO) has a critical role in angiogenesis, the therapeutic potential of NO synthase overexpression in severe ischemia remains undefined. We tested the hypothesis that overexpression of endothelial NO synthase (eNOS) would improve tissue perfusion in severe hind limb ischemia.Methods: Severe hind limb ischemia was induced in 122 adult male Sprague-Dawley rats. Ten days after the induction of hind limb ischemia, vascular isolation and intraarterial delivery of an adenoviral vector encoding eNOS (AdeNOS), a control adenoviral vector (AdE1), or phosphate-buffered saline solution (PBS) was performed. Skeletal muscle blood flow, muscle oxygen tension, angiography, and immunohistochemistry for capillary counts were measured.Results: Gene transfer of AdeNOS increased eNOS protein expression and enzyme activity. Two weeks after gene transfer, skeletal muscle blood flow was fourfold higher in eNOS-transduced than in AdE1-transduced or PBS treated rats and was similar to exercise-induced maximal flow in nonischemic muscle. eNOS overexpression increased muscle oxygen tension in a titer-dependent fashion. This increase persisted 1 month after transduction, even though eNOS enzyme activity had declined to normal levels. Angiography and capillary counts showed that eNOS overexpression increased the size and number of collateral arteries, but did not significantly increase the capillary muscle fiber ratio.Conclusions: eNOS overexpression in an ischemic rat hind limb significantly increased skeletal muscle blood flow, muscle oxygen tension, and collateral arteries (arteriogenesis). Furthermore, eNOS overexpression did not result in capillary angiogenesis above control levels. These studies demonstrate the potential for eNOS overexpression as treatment for severe limb ischemia in human beings.