CD151 promotes neovascularization and improves blood perfusion in a rat hind-limb ischemia model
CD151 promotes neovascularization and improves blood perfusion in a rat hind-limb ischemia model
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DOI：
10.1583/04-1478r.1
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发表时间：
2005-08-01
影响因子：
2.6
通讯作者：
Wang, DW
中科院分区：
文献类型：
--
作者：
Lan, RF;Liu, ZX;Wang, DW
目的：评价重组腺相关病毒（rAAV）介导的CD151基因递送促进大鼠后肢缺血模型骨骼肌新生血管形成和改善血液灌注的效率。方法：将CD151构建到rAAV载体中。将12只Wistar大鼠随机分为2组，每组6只，然后在一侧后肢分别肌肉注射rAAV-CD151或rAAV-GFP。基因递送两周后，切除治疗肢体的股动脉以建立后肢缺血模型。通过蛋白质印迹和逆转录聚合酶链式反应证实转基因产物CD151的表达。转染后6周，测量后肢的皮肤温度、血管造影评分和毛细血管密度，以评估血液灌注和新生血管形成。结果：与转染GFP的组相比，CD151组的血管造影评分高出63%（p < 0.05），毛细血管密度增加18%（p < 0.05）。此外，AAV-CD151转染后肢的平均皮肤温度与对侧非缺血肢体的水平相当，而GFP转染大鼠的肢体温度显着低于非缺血对照。注射rAAV-CD151的缺血后肢中CD151的表达显着高于注射rAAV-GFP的肢体。注射后至少 6 周，在注射肌肉中持续观察到 CD151 mRNA 和蛋白表达，而在远端器官或注射 rAAV-GFP 的肌肉中未检测到人 CD151 mRNA。 结论：本研究表明，rAAV 介导的 CD151 基因转移到大鼠骨骼肌中是有效、稳定的，并且没有异位表达。此外，rAAV介导的CD151基因转移刺激新血管形成，特别是动脉生成，从而改善大鼠后肢缺血模型的血液灌注。这些发现表明CD151可能成为缺血性疾病新生血管治疗的新靶点，rAAV介导的CD151基因转移可能有助于治疗缺血性疾病。
Purpose: To evaluate the efficiency of recombinant adeno-associated virus (rAAV)-mediated CD151 gene delivery in promoting neovascularizalion and improving blood perfusion in the skeletal muscle of the rat hind-limb ischemia model.Methods: CD151 was constructed into the rAAV vector. Twelve Wistar rats were randomly divided into 2 groups of 6 rats each and then intramuscularly injected with rAAV-CD151 or rAAV-GFP, respectively, in one hind limb. Two weeks after gene delivery, the femoral arteries in the treated limbs were excised to establish the model of hind-limb ischemia. Expression of the transgene product CD151 was confirmed by Western blot and the reverse transcription polymerase chain reaction. The skin temperature, angiographic score, and capillary density of the hind limb were measured to assess blood perfusion and neovascularization 6 weeks after transfection.Results: Compared to the group transfected with GFP, the CD151 group showed a 63% higher angiographic score (p < 0.05) and an 18% increase in capillary density (p < 0.05). In addition, the mean skin temperature of the AAV-CD151-transfected hind limbs was equivalent to the level of the contralateral nonischemic limb, whereas the limb temperature in the GFP-transfected rats was significantly lower than the nonischemic control. The expression of CD151 in the ischemic hind limb injected with rAAV-CD151 was significantly higher than limbs injected with rAAV-GFP. The CD151 mRNA and protein expression was persistently observed in the injected muscle for at least 6 weeks after injection, while no human CD151 mRNA could be detected in remote organs or rAAV-GFP-injected muscles.Conclusions: This study demonstrates that the rAAV-mediated CD151 gene transfer into rat skeletal muscles is efficient, stable, and has no ectopic expression. Moreover, rAAV-mediated CD151 gene transfer stimulates neovascularization, especially arteriogenesis, and thereby improves blood perfusion in a rat hind-limb ischemia model. These findings suggest that CD151 could be a new target for neovascularization therapy in ischemic disease, and rAAV-mediated CD151 gene transfer may be useful for treatment of ischemic disease.