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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。
AIM: To investigate the role of muscle-derived recombinant adeno-associated virus (rAAV) -mediated CD151 gene in proangiogenesis and revascularization in rat hindlimb ischemia model. Methods: Recombinant adeno-associated virus (rAAV-CD151, rAAV-GFP) carrying CD151 gene and GFP gene were packaged respectively. Twelve Wistar rats were randomly divided into experimental group and control group with 6 rats in each group. Rats in the experimental group were transfused with rAAV-CD151 intramuscularly and control group with rAAV-GFP. The left hindlimb ischemia model was established by femoral artery excision two weeks after transfection. Vascular remodeling of ischemic limbs and evaluation of revascularization were observed by hind limb angiography and capillary density of ischemic muscle tissue. Western blot was used to detect CD151 expression in ischemic limbs in both groups. Results: After 4 weeks of femoral artery resection (6 weeks of gene transfection), the results of hind limb angiography showed that the mean ischemic vessel score was 2.56 ± 0.37 in the experimental group and 1.57 ± 0.39 in the control group. The ischemic hind limb muscle tissue The capillary density was (609.00 ± 83.89) / mm2, while the control group was (517.00 ± 70.65) / mm2. There was significant difference between the two groups (P <0.05). The results of Western blot showed that the expression of CD151 in ischemic muscle of experimental group was 2.96 times of the control group. CONCLUSION: Local high expression of CD151 gene can improve the ability of revascularization of ischemic tissue and promote the revascularization of ischemic tissue. CD151 will provide a new target for angiogenesis of ischemic disease.