【摘要】 目的 :RNA干扰 (RNAi)是一种新的高效特异地阻断基因表达的基因阻断技术。本研究旨在探讨血管紧张素Ⅱ 1型受体 (AT1R)的短发夹环RNA质粒 (pAT1R shRNA)对大鼠血管平滑肌细胞 (VSMC)增殖的影响。 方法 :构建pAT1R shRNA质粒 ,并转染入鼠VSMC中 ,应用RT PCR及Westernblot检测血管平滑肌细胞AT1R的mRNA和蛋白表达的变化 ,验证 pAT1R shRNA是否具有RNAi作用 ;应用台盼蓝染色法和MTT法检测VSMC增殖情况。结果 :构建的质粒 pAT1R shRNA经测序鉴定验证与预期相符。转染组AT1R的mRNA和蛋白表达明显减少 ,与对照组相比有显著性差异 (P <0 .0 1) ;转染质粒同时加AngⅡ刺激的VSMC增生明显受到抑制 ,与对照组相比有显著性差异 (P <0 .0 1)。结论 :构建的pAT1R shRNA质粒具有RNAi作用 ,并对AngⅡ所诱导的VSMC增殖有抑制作用更多还原

【Abstract】 Aim: RNA interference is a new technology that inhibit effectively of the expression the specific genes. The present study was designed to investigate whether the plasmid containing the short hairpin RNA(shRNA) of angiotensin Ⅱ type 1 receptor(AT1R) can inhibit the hyperplasia of VSMCs in rat. Methods: The plasmids containing the shRNA of AT1R were constructed, and transfected vascular smooth musle cell(VSMC) to detect the effect on the AT1R expression by RT-PCR and Western blot, and detect the hyperplasia of VSMCs by trypan blues taining and MTT. Results: The plasmids was certified to be in the right rank, and there was significant difference (P<0.01) in the expression of AT1R mRNA and protein between the plasmid transfected group and the control group. There was significant difference(P<0.01) in the hyperplasia of VSMCs between the plasmid transfected adding AngⅡ group and the control group. Conclusion: The plasmids containing the shRNA of AT1R have the effect of RNAi, and inhibit the hyperplasia of VSMCs induced by AngⅡ in rat.更多还原