【摘要】 目的　探讨纤溶酶原激活物抑制剂 - 1(plasminogen activator inhibitor- 1,PAI- 1)反义 RNA对离体培养的主动脉平滑肌细胞 (smooth muscle cell,SMC) PAI- 1表达的作用及对血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的影响。方法　PCR扩增 PAI- 1第 2外显子 ,将 PCR产物纯化克隆后连入真核细胞表达载体 pc DNA3.1,构建 PAI- 1反义 RNA重组质粒。将 pc DNA3.1-反义PAI- 1重组质粒转染 SMC中。通过免疫组化、Western印迹、EL ISA检测细胞中 PAI- 1表达的改变 ;通过免疫荧光技术观察细胞中 PAI- 1表达量的变化对 VEGF的影响。结果　转染后第 3天 ,细胞中 PAI- 1含量最低 ,VEGF的表达也减少。第 5天 ,PAI- 1含量逐渐升高 ,VEGF也相应增加。第 7天 ,PAI- 1含量接近于正常 ,VEGF也增至正常水平。结论　反义 PAI- 1RNA能有效阻断 SMC中 PAI- 1的蛋白合成 ,同时抑制细胞中 VEGF的表达。更多还原

【Abstract】 Objective To investigate the role of plasminogen activator inhibitor 1(PAI 1) antisense RNA in regulating the expression of PAI 1 and vascular endothelial growth factor(VEGF) in aorta smooth muscle cells cultured in vitro. Methods The second extron of PAI 1 was amplified with polymerase chain reaction(PCR), the production was inserted into eukaryotic cell expression vector pcDNA3.1 after it had been purified and cloned so as to construct PAI 1 antisence RNA recombination plasmid. The recombination plasmid was transfected into SMC. PAI 1 expression was detected by immumohistochemistry, Western blod and ELISA; the effects of PAI 1 variation on VEGF was examined by immunofluorescence. Results PAI 1 antigen was the lowest in cells on the third day after transfection; the expression of VEGF was also decreased. PAI 1 antigen gradually increased on the fifth day and VEGF increased correspondently. On the seventh day, PAI 1 antigen and VEGF increased to nearly normal level. Conclusion PAI 1 antisense RNA can block the translation progress of PAI 1 proteins effectively and inhibit the expression of VEGF in aorta smooth muscle cells.更多还原