【摘要】 目的:研究聚四氟乙烯(PTFE)血管材料携载血管内皮生长因子(VEGF)基因并促进内皮细胞生长的可行性.方法:将pCDI-hVEGF121/pCDI质粒溶液处理过的PTFE人工血管修剪成圆片,置于生理盐水中并于0.5,1,2,4,8,16,24,48和72h测定溶液的吸光度,计算出溶液的DNA浓度,绘出体外释放曲线.分离和培养人脐静脉内皮细胞,将内皮细胞种植到携带基因质粒的PTFE材料表面,在6,24,72和120h检测细胞增殖情况并用ELISA法检测培养液中VEGF蛋白的表达量.结果:体外释放曲线提示在0.5~4h的这段时间内,质粒释放较快,随后进入缓慢增长期,直至72h仍有质粒的释放.在24,72和120h时间点,VEGF质粒组PTFE材料上的内皮细胞数和增长倍率都明显高于空质粒组材料(P<0.05),在6,24,72和120h时间点,VEGF质粒组培养液的VEGF蛋白水平和增长倍率也明显高于空质粒组(P<0.01).结论:证实了PTFE材料携带VEGF基因转染内皮细胞并具有促进其生长的可能性.更多还原

【Abstract】 AIM: To investigate whether vascular endothelial growth factor (VEGF) gene plasmid carried by polytetrafluoroethylene (PTFE) vascular grafts can be transfered to endothelial cells (ECs) and promote the growth of ECs. METHODS: The PTFE vascular grafts carrying pCDI-hVEGF_ 121 or pCDI were soaked in normal saline. The optical density of the solution at 260 nm and the DNA concentration were measured at 0.5, 1, 2, 4, 8, 16, 24, 48 and 72 h, then the release curve was made. ECs were derived from human umbilical vein. ECs were seeded on the PTFE materials carrying pCDI-hVEGF121 or pCDI, the number of ECs was counted and VEGF protein concentration was measured by ELISA method at 6, 24, 72 and 120 h. RESULTS: The release curve showed the plasmid was released quickly during 0.5- 4 h, then slowly, but continuously even after 72 h. At 24, 72 and 120 h, the number and proliferation rate of ECs on PTFE materials carrying pCDI-hVEGF121 were higher than those on PTFE materials carrying pCDI (P<0.05). VEGF protein concentration of the former was higher than that of the latter at 6, 24, 72 and 120 h (P<0.01). CONCLUSION: The PTFE graft can be used as a carrier of VEGF gene plasmid and help VEGF gene transfection into ECs for promoting the growth of ECs.更多还原