Yamada Narihisa, Li Wei, Ihaya Akio, Kimura Tetsuya, Morioka Kouichi, Uesaka Takahiko, Takamori Atsushi, Handa Mitsuteru, Tanabe Sawaka, Tanaka Kuniyoshi
Second Department of Surgery, Faculty of Medical Sciences, University of Fukui, Fukui, Japan.
J Vasc Surg. 2006 Dec;44(6):1322-8. doi: 10.1016/j.jvs.2006.07.051.
Platelet-derived endothelial cell growth factor (PD-ECGF) is identical to thymidine phosphorylase (TP), and it can induce angiogenesis, including arteriogenesis, in chronically ischemic canine myocardium. Because its effect on peripheral arterial disease has not been elucidated, we investigated whether overexpression of PD-ECGF/TP could ameliorate chronic limb ischemia in rabbits.
Left femoral arteries were resected from 24 male rabbits. After 10 days, a plasmid vector containing human PD-ECGF/TP complimentary DNA was injected into 10 sites in the adductor muscles. Control groups received either the LacZ plasmid vector or saline vehicle only (n = 8 per group). Blood pressure was measured in the calf before surgery, at the onset of ischemia, 10 days later, and 20 and 30 days after gene transfer. Collateral vessel development and limb perfusion were assessed by angiography, and resected tissues underwent molecular and histologic examination.
In the PD-ECGF/TP group, human PD-ECGF/TP messenger RNA and protein were still detected at 30 days after treatment. Calf blood pressure decreased significantly after femoral artery resection in all three groups. It subsequently showed a greater increase in the PD-ECGF/TP group than in either control group, and the difference was significant at 20 days after treatment (PD-ECGF/TP, 97.4 +/- 7.4; LacZ, 58.6 +/- 6.9; saline, 41.3 +/- 3.6). Immunohistochemical staining demonstrated an increased ratio of capillaries and arterioles to muscle fibers in the PD-ECGF/TP group (2.14 +/- 0.13 and 1.51 +/- 0.06), but not in the LacZ group (1.39 +/- 0.04 and 0.71 +/- 0.05) or the saline group (1.34 +/- 0.05 and 0.71 +/- 0.04, P < .01). The angiographic score was higher in the PD-ECGF/TP group (0.96 +/- 0.08) than in the LacZ group (0.50 +/- 0.02) or saline group (0.51 +/- 0.03) at 30 days after gene transfer (P < .01).
This study demonstrated that PD-ECGF/TP gene transfer induced angiogenesis and decreased ischemia in a rabbit hindlimb model by promoting arteriogenesis, suggesting that targeting this gene may be a promising therapeutic strategy for peripheral vascular disease.
血小板衍生内皮细胞生长因子（PD-ECGF）与胸苷磷酸化酶（TP）相同，它能够在慢性缺血的犬心肌中诱导血管生成，包括动脉生成。由于其对外周动脉疾病的影响尚未阐明，我们研究了PD-ECGF/TP的过表达是否能改善兔慢性肢体缺血。
从24只雄性兔身上切除左股动脉。10天后，将含有人类PD-ECGF/TP互补DNA的质粒载体注射到内收肌的10个部位。对照组分别接受LacZ质粒载体或仅接受生理盐水（每组n = 8）。在手术前、缺血开始时、10天后以及基因转移后20天和30天测量小腿血压。通过血管造影评估侧支血管的发育和肢体灌注，并对切除的组织进行分子和组织学检查。
在PD-ECGF/TP组中，治疗后30天仍可检测到人类PD-ECGF/TP信使核糖核酸和蛋白质。在所有三组中，股动脉切除后小腿血压均显著下降。随后，PD-ECGF/TP组的血压升高幅度大于任何一个对照组，且在治疗后20天差异显著（PD-ECGF/TP组，97.4±7.4；LacZ组，58.6±6.9；生理盐水组，41.3±3.6）。免疫组织化学染色显示，PD-ECGF/TP组中毛细血管和小动脉与肌纤维的比例增加（分别为2.14±0.13和1.51±0.06），而LacZ组（分别为1.39±0.04和0.71±0.05）和生理盐水组（分别为1.34±0.05和0.71±0.04，P <.01）则未增加。基因转移后30天，PD-ECGF/TP组的血管造影评分（0.96±0.08）高于LacZ组（0.50±0.02）和生理盐水组（0.51±0.03）（P <.01）。
本研究表明，PD-ECGF/TP基因转移通过促进动脉生成在兔后肢模型中诱导血管生成并减轻缺血，提示靶向该基因可能是外周血管疾病的一种有前景的治疗策略。
