In mice with type 2 diabetes, a vascular endothelial growth factor (VEGF)-activating transcription factor modulates VEGF signaling and induces therapeutic angiogenesis after hindlimb ischemia
In mice with type 2 diabetes, a vascular endothelial growth factor (VEGF)-activating transcription factor modulates VEGF signaling and induces therapeutic angiogenesis after hindlimb ischemia
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DOI：
10.2337/db06-0999
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发表时间：
2007-03-01
期刊：
影响因子：
7.7
通讯作者：
Annex, Brian H.
中科院分区：
文献类型：
--
作者：
Li, Yongjun;Hazarika, Surovi;Annex, Brian H.
外周动脉疾病是糖尿病的主要并发症。促进治疗性血管生成的能力在糖尿病患者中可能是有限的。用高脂饲料喂养C57BL/6小鼠(n=60)建立2型糖尿病模型。正常饲料喂养的小鼠(n=20)没有糖尿病。小鼠接受单侧股动脉结扎和切除。一种编码旨在增加血管内皮生长因子表达的工程转录因子(ZFP-VEGF)的质粒DNA。术后第10天，缺血侧肢体给予125mgZFP-VEGF质粒或空白对照。分别于注射后第3、10、20天处死小鼠(n=10/组，各时间点)。激光多普勒血流成像检测肢体血流量。实时定量聚合酶链式反应检测血管内皮细胞生长因子基因的表达。采用酶联免疫吸附试验检测血管内皮细胞生长因子、Akt和磷酸化Akt蛋白。组织学检测毛细血管密度、细胞增殖和细胞凋亡。与正常小鼠相比，糖尿病小鼠的血管内皮生长因子蛋白表达增加，结扎前磷酸化Akt/Akt比率降低，结扎后血流灌注恢复受损。在注射后3天和10天，糖尿病小鼠的基因转移增加。血管内皮生长因子的表达和信号转导。在以后的时间点，基因转移导致更好的血流灌注恢复。ZFP-VEGF基因转移可促进2型糖尿病小鼠治疗性血管生成。
Peripheral arterial disease is a major complication of diabetes. The ability to promote therapeutic angiogenesis may be limited in diabetes. Type 2 diabeters was induced by high-fat feeding C57BL/6 mice (n = 60). Normal chow-fed mice (n = 20) had no diabetes. Mice underwent unilateral femoral artery ligation and excision. A plasmid DNA encoded an engineered transcription factor designed to increase vascular endothelial growth factor expression (ZFP-VEGF). On day 10 after the operation, the ischemic limbs received 125 mu g ZFP-VEGF plasmid or control. Mice were killed 3, 10, or 20 days after injection (n = 10/group, at each time point). Limb blood flow was measured by laser Doppler perfusion imaging. VEGF mRNA expression was examined by real-time PCR. VEGF, Akt, and phospho-Akt protein were measured by enzyme-linked immunosorbent assay. Capillary density, proliferation, and apoptosis were assessed histologically. Compared with normal mice, mice with diabetes had greater VEGF protein, reduced phospho-Akt-to-Akt ratio before ligation, and an impaired perfusion recovery after ligation. At 3 and 10 days after injection, in mice with diabetes, gene transfer increased. VEGF expression and signaling. At later time points, gene transfer resulted in better perfusion recovery. Gene transfer with ZFP-VEGF was able to promote therapeutic angiogenesis mice with type 2 diabetes.