腺病毒介导的VEGF-B基因促血管内皮细胞增殖作用的研究

为观察腺癌毒素介导的血管内皮细胞生长因子B（VEGF－B）基因体外转染对血管内皮细胞的促增殖作用,构建编码人VEGF－B基因的复制缺陷的重组腺病毒载体,体外转染鼠主动脉血管内皮细胞（RAECs）,应用RT－PCR和Western blot检测外源VEGF－B的表达,应用四唑盐（MTT）观察转染后RAECs的增殖。结果发现,RAECs可有效地被重组腺病毒载体感染,并能成功转录和表达VEGF－B基因和蛋白,在转染后细胞培养上清中检测到VEGF－B蛋白的表达,对RAECs有显著促增殖作用。提示重组腺病毒载体介导的人VEGF－B基因有血管新生作用,可用于缺血性心脏病的治疗。

ADENOVIRUS-MEDIATED GENE TRANSFER OF VASCULAR ENDOTHELIAL GROWTH FACTOR B INDUCES PROLIFERATION OF VASCULAR ENDOTHELIAL CELLS

To investigate the effect of transfection of adenovirus mediated vascular endothelial growth factor B (VEGF B) gene transfer on the proliferation of endothelial cells. We constructed a replication deficient recombinant adenovirus vector coding for human VEGF B, and detected its gene transcription and expression in Ad. VEGF B infected rat arotic endothelial cells (RAECs) with RT PCR and western blat. The effect of Ad.VEGF B on RAECs prolifeation was examined by MTT assay on days 1, 2, 3, 5, 7 after infection. RAECs were successfully infected with high efficiency and expressed a special protein that is immunoreactive to anti VEGF B monoclonal antibody. This protein could be detected in the culture medium. The proliferation assay of RAECs infected with VEGF B expressing adenovirus revealed a significant increase in cell number over control. These results implied that the use of VEGF B expressing recombinant adenovirus may induce angiogenesis, which might be potential in vivo gene therapy for ischemic heart disease.