| 携载VEGF基因的聚四氟乙烯血管材料对内皮细胞生长的促进作用
| | 药学本科毕业论文 作者：陶思丰，陈力，许远，田华，李国刚，陈健
【关键词】 聚四氟乙烯；血管内皮生长因子类；人工血管；基因；内皮,血管/细胞学
【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 pCDIhVEGF121 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 pCDIhVEGF121 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 pCDIhVEGF121 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.
【Keywords】 polytetrafluoroethylene; vascular endothelial growth factors; blood vessel prosthesis; genes; endothelium, vascular/cytology
【摘要】 目的药学本科毕业论文： 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药学本科毕业论文可行性. 方法： 将pCDIhVEGF121/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72 h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120 h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0.5~4 h的这段时间内，质粒释放较快，随后进入缓慢增长期，直至72 h仍有质粒的释放. 在24, 72和120 h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120 h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.
【关键词】 聚四氟乙烯；血管内皮生长因子类；人工血管；基因；内皮，血管/细胞学
0引言
聚四氟乙烯（polytetrafluoroethylene, PTFE）作为良好的人工血管材料而广泛应用于临床，但如何促进移植血管表面内皮细胞的生长而提高通畅率还是一个有待研究的课题. 近年来利用基因治疗血管性疾病已经取得了一定的疗效，并开始运用基因来改造人工血管以期获得促进内皮生长的效果. 在众多已发现的诱导血管生长的因子中，血管内皮生长因子(vascular endothelial growth factor, VEGF)被认为是诱导血管内皮细胞生长作用最强、特异性最高的一种生长因子［1-2］. 我们利用PTFE自身多孔的特性来携带VEGF基因，并将血管内皮细胞种植到其表面，期望VEGF基因能够转染内皮细胞并促进其生长. 本实验就是基于上述设想，探讨了PTFE材料携带VEGF基因并转染内皮细胞的可能性.
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