rAAV-CD151基因在大鼠缺血后肢血管再生中的作用及血管造影评分

Effects of rAAV-CD151 Gene on the Neovascularization and Angioscore of Angiography in the Model of Rat Hind-limb Ischemia

目的:研究肌肉转染重组腺相关病毒(recombinant adeno-associated virus,rAAV)介导的CD151基因在大鼠后肢缺血模型中促血管再生和血运重建的作用。方法:分别包装携带有CD151基因和GFP基因的重组腺相关病毒(rAAV-CD151,rAAV-GFP)。Wistar大鼠12只,随机分为实验组和对照组,每组各6只。实验组大鼠左下肢肌肉注射局部转染rAAV-CD151,对照组注射rAAV-GFP,基因转染2周后行股动脉切除术建立左侧大鼠后肢缺血模型。通过后肢血管造影和缺血肌肉组织毛细血管密度检测,观察缺血肢体血管再生和评价血运重建情况。Western blot检测两组大鼠局部缺血肢体CD151表达。结果:股动脉切除术4周(基因转染6周)后,后肢血管造影结果显示实验组缺血侧血管评分平均为2.56±0.37,对照组平均为1.57±0.39;实验组缺血后肢肌肉组织毛细血管密度为(609.00±83.89)/mm2,对照组为(517.00±70.65)/mm2;两组比较差异均有显著性意义(P<0.05)。Western blot检测结果显示实验组大鼠缺血侧肌肉组织CD151表达为对照组的2.96倍。结论:局部高表达CD151基因能够提高缺血组织的血管再生能力,促进缺血组织的血运重建;CD151将为缺血性疾病的血管再生治疗提供新的靶点。

Objective：To investigate the effects of AAV-mediated CD151 gene on neovascularization and the angioseore of angiography in the model of rat hind-limb ischemia. Methods：The recombinant adeno-associated virus vector containing CD151 gene （rAAV-CD151） and the recombinant adeno-associated virus vector containing sense GFP gene （rAAV- GFP） were produced. 12 Wister rats were randomly divided into two groups. The rats of experimental group and control group were intramuscularly injected rAAV CD151 and rAAV-GFP （1 × 1010pfu/rat） respectively. After 2 weeks of transfection . the femoral arteries of the, rats were excised to establish the model of left hind-limb ischimia. The contrast angiography of the hind-limb and the capillary density of the ischemic muscular were detected to measure the formation of new blood vessels. The expression of the CD151 in the ischemic tissue was detected by Western blot. Results：The hind-limb angiography showed that the average angiographic score was 2.56±0.37 in experimental group and 1.57±0.39 in control group. The capillary density was 609 ±83.89/mm^2 in experimental group and 517 ± 70.65/mm^2 in control group. All the differences between the two groups were statistically significant （P〈0.05）. The expression of CD151 in ischemic hind-limb of experimental group was 2.96 times as high as control group. Conclusion： High expression of CD151 in the ischemic tissue can stimulate the formation of the new blood vessels and promote the foundation of the blood flow. The CD151 may be a new target of the neovascutarization therapy in the ischemic disease.