Rescue of diabetes-related impairment of angiogenesis by intramuscular gene therapy with adeno-VEGF
Rescue of diabetes-related impairment of angiogenesis by intramuscular gene therapy with adeno-VEGF
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DOI：
10.1016/s0002-9440(10)65282-0
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发表时间：
1999-02-01
影响因子：
6
通讯作者：
Isner, JM
中科院分区：
文献类型：
--
作者：
Rivard, A;Silver, M;Isner, JM
糖尿病是冠状动脉和外周动脉疾病的主要危险因素。尽管糖尿病患者经常出现这些疾病的晚期形式，但尚不清楚血管缺血的代偿机制是否在这种情况下受到影响。因此，我们试图确定糖尿病是否可以：1）损害响应组织缺血的新侧支血管形成的发展，2）抑制细胞因子诱导的治疗性新血管形成，后肢缺血是通过非肥胖糖尿病小鼠（NOD小鼠，n = 20）和对照C57小鼠（n = 20）的股动脉结扎造成的。手术后通过连续激光多普勒研究评估后肢灌注。在 NOD 小鼠中，对缺血肢体和正常肢体之间的多普勒弓比的测量表明，缺血后肢的灌注恢复明显受损。手术后第 14 天，NOD 小鼠的多普勒血流比率为 0.49 +/- 0.04，而 C57 小鼠为 0.73 +/- 0.06（P 小于或等于 0.005）。这种血弓恢复损伤在整个研究期间持续存在，第 35 天的多普勒血流比率值为 0.50 +/- 0.05 与 0.90 +/- 0.07分别在 NOD 和 C57 小鼠中（P 小于或等于 0.001），CD31 免疫染色证实了激光多普勒数据，显示手术后 35 天 NOD 小鼠中毛细血管密度显着降低（302 +/- 4 毛细血管/mm(2)，而 C57 小鼠中为 782 +/- 78（P 小于或等于 0.005）。NOD 小鼠中新生血管的减少是缺血组织中血管内皮生长因子 (VEGF) 水平较低的结果，通过 Northern 印迹、蛋白质印迹和免疫组织化学评估，通过显示通过肌内注射编码 VEGF 的腺病毒载体补充该生长因子的 NOD 小鼠可以达到正常水平的新血管形成，证实了 VEGF 的核心作用。缺血组织； 2) VEGF表达减少导致新血管形成受损； 3）通过肌内腺-VEGF基因转移实现的细胞因子补充可恢复糖尿病小鼠模型中的新血管形成。
Diabetes is a major risk factor for coronary and peripheral artery diseases. Although diabetic patients often present with advanced forms of these diseases, it is not known whether the compensatory mechanisms to vascular ischemia are affected in this condition. Accordingly, we sought to determine whether diabetes could: 1) impair the development of new collateral vessel formation in response to tissue ischemia and 2) inhibit cytokine-induced therapeutic neovascularization, Hindlimb ischemia was created by femoral artery ligation in nonobese diabetic mice (NOD mice, n = 20) and in control C57 mice (n = 20). Hindlimb perfusion was evaluated by serial laser Doppler studies after the surgery. In NOD mice, measurement of the Doppler Bow ratio between the ischemic and the normal limb indicated that restoration of perfusion in the ischemic hindlimb was significantly impaired. At day 14 after surgery, Doppler flow ratio in the NOD mice was 0.49 +/- 0.04 versus 0.73 +/- 0.06 for the C57 mice (P less than or equal to 0.005), This impairment in blood Bow recovery persisted throughout the duration of the study with Doppler Bow ratio values at day 35 of 0.50 +/- 0.05 versus 0.90 +/- 0.07 in the NOD and C57 mice, respectively (P less than or equal to 0.001), CD31 immunostaining confirmed the laser Doppler data by showing a significant reduction in capillary density in the NOD mice at 35 days after surgery (302 +/- 4 capillaries/mm(2) versus 782 +/- 78 in C57 mice (P less than or equal to 0.005). The reduction in neovascularization in the NOD mice was the result of a lower level of vascular endothelial growth factor (VEGF) in the ischemic tissues, as assessed by Northern blot, Western blot and immunohistochemistry. The central role of VEGF was confirmed by showing that normal levels of neovascularization (compared with C57) could be achieved in NOD mice that had been supplemented for this growth factor via intramuscular injection of an adenoviral vector encoding for VEGF. We conclude that 1) diabetes impairs endogenous neovascularization of ischemic tissues; 2) the impairment in new blood vessel formation results from reduced expression of VEGF; and 3) cytokine supplementation achieved by intramuscular adeno-VEGF gene transfer restores neovascularization in a mouse model of diabetes.