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脂质体介导的VEGF基因对培养血管细胞增殖的影响 |
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蒋学俊 魏炜 熊一力 陆再英
摘要 目的：观察脂质体介导转移的VEGF基因在血管平滑肌细胞（VSMC）中的表达，比较VEGF对血管内皮细胞（VEC）及VSMC增殖的影响。方法：将含血管内皮生长因子（VEGF）cDNA的真核表达载体pSVI21用脂质体介导的基因转移法导入血管平滑肌细胞（VSMC）中，取此VSMC条件培养液，再行血管内皮细胞（VEC）培养，用Northern blot杂交，Western印迹法及［3H］胸腺嘧啶核苷掺入法，观察了VEGF在VSMC及其条件培养液中的表达，比较了VEGF对VEC及VSMC增殖的影响。结果：转基因组VSMC中VEGF mRNA 呈稳定高表达，转基因组VSMC条件培养液中，VEGF抗原表达显著高于对照组（P均＜0.01），加入此条件培养液的各组VEC的［3H］胸腺嘧啶核苷掺入量均显著高于对照组（P均＜0.01），而在VSMC组无显著差异。结论：VEGF的转录和表达表明脂质体介导的血管细胞VEGF基因转移是成功的。VEGF具有促VEC增殖作用，但对VSMC无促增殖作用，有利于缺血性疾病及血管再狭窄的防治。
主题词 内皮生长因子；脂质体；转染；肌，平滑
Effect of liposome-mediated vascular endothelial growth factor gene
on proliferation of cultured vascular cells
JIANG Xue-Jun,WEI Wei,XIONG Yi-Li,LU Zai-Ying
Department of Cardiology,Affiliated Tongji Hospital,Tongji Medical University,Wuhan (430030)
Abstract AIM: Observe the expression of liposome-mediated vascular endothelial growth factor (VEGF) gene of in vitro vascular smooth muscle cells(VSMC),and compare the effect of VEGF on proliferation of vessel endothelium cell and VSMC.METHODS: An eukaryotic expression vector pSVI21 containing VEGF cDNA was transferred into VSMC in vitro by lipofectamineTM reagent-mediated method.Vascular endothelial cells(VEC) were cultured with the above VSMC conditioned medium.In order to determine the expression of VEGF mRNA in VSMC and VEGF antigen in VSMC conditional medium,and compared the effect of VEGF on proliferation of VEC and VSMC,Northern blot,Western blot and ［3 H］-thymidine (［3 H］TdR) incorporation were used.RESULTS: The expression of VEGF mRNA in transgene VSMC groups was higher.The expression of VEGF antigen in transgenic medium was significantly higher (P＜0.01) than control group,［3 H］TdR data(counts.min-1) was significantly higher in VEC groups added transgenic medium than control group(P＜0.01),but there was no significant difference between VSMC groups.CONCLUSION: The method of liposome-mediated VEGF gene transfer into VSMC is successf[1] [2] 下一页
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