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(1).docxVIP

携载VEG嘎因的聚四氟乙烯血管材料对内 皮细胞生长的促进作用（1） 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 gene plasmid carried by polytetrafluoroethylene vascular grafts can be transfered to endothelial cells and promote the growth of ECs. METHODSThe PTFE vascular grafts carrying pCDIhVEGF121orpCDI were soaked in normal saline. The optical density of the solution at 260 nm and the DNA concentration were measured at , 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 pCDIhVEGF121orpCDI, 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 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 pCDIhVEGF121were higher than those on PTFE materials carrying pCDI . VEGF protein concentration of the former was higher than that of the latter at 6, 24, 72 and 120 h . CONCLUSION: The PTFE graft can be used as a carrier of VEGFgene plasmid and help VEGFgene 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：研究聚四氟乙烯血管材料携载血管内 皮生长因子基因并促进内皮细胞生长的可行性 .方法： 将 pCDIhVEGF121/pCDI质粒溶液处理过的 PTFEA工血管修剪成 圆片，置于生理盐水中并于 ,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＜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 表达虽.结果： 体外释放曲线提示在~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，VEG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， 在6, 24, 72和120 h时间点，VEG质粒组培养液的 VEGF?白水 平和增长倍率也明显高于空质粒组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 VEG建因，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