体外培养人脑动静脉畸形血管内皮细胞中VEGF表达增加

Detection of VEGF Expression in Cultured Human Cerebral Arteriovenous Malformation Vascular Endothelial Cells in Vitro

目的探讨体外培养的人脑动静脉畸形血管内皮细胞和正常脑血管内皮细胞中血管内皮细胞生长因子(VEGF)表达水平的差异。方法采用组织块贴壁法对人脑动静脉畸形血管内皮细胞进行培养和形态学观察;免疫组化检测CD31和vWF抗原验证内皮细胞;采用RT-PCR和Western blot技术检测脑动静脉畸形血管内皮细胞VEGF mRNA和蛋白表达,并与和正常脑血管内皮细胞进行比较。结果体外培养的内皮细胞CD31和vWF染色阳性率达95%以上。人脑动静脉畸形血管内皮细胞中VEGF mRNA和蛋白表达均显著升高(P<0.05)。结论体外培养的人脑动静脉畸形内皮细胞中VEGF的高表达,提示血管生成在脑动静脉畸形的发病机制中起重要作用。

Aim To compare the expression level of vascular endothelial growth factor （VEGF） in human cerebral arteriovenous malformation endothelial cells （AVMEC） and normal brain endothelial cell （BECs） in vitro. Methods The endothelial cells were isolated and cultured by the vascular tissue explants adherent method. Endothelial cells markers CD31 and vWF were immunohistochemically detected. Expression levels of VEGF mRNA and protein were tested by RT-PCR and Western blot, and were compared. Results More than 95% of the cells were positive for CD31 and vWF staining. Expression levels of VEGF mRNA and protein were significantly higher in AVMEC than in BECs （P〈0.05）. Conclusion Cultured AVMEC expressed higher VEGF than the controlled BEC, implicating angiogenesis played an important role in the pathogenesis of cerebral arteriovenous malformations.