摘要
目的:研究腺病毒介导反义AT1R cDNA转染对人肺动脉平滑肌细胞(PASMCs)增殖的影响。方法:构建重组反义人AT1R腺病毒(AdCMVahAT1)和表达β-半乳糖苷酶的对照载体AdCMVLacZ,将培养的PASMCs分为DMEM组、ATⅡ组、AdCMVLacZ+ATⅡ组和AdCMVahAT1+ATⅡ组,分别给予相应的干预因素,用RT-PCR和免疫组化检测AT1R的表达,用流式细胞仪检测各组PASMCs的增殖指数。结果:转染病毒后48 h AT1R蛋白表达在AdCMVahAT1组显著低于AdCMVLacZ组和DMEM组。给予ATⅡ刺激48 h,ATⅡ组PASMCs的增殖指数(59.69±3.46)高于DMEM组(50.25±1.34,P<0.01),转染AdCMVahAT1后再用ATⅡ刺激48 h的AdCMVahAT1+ATⅡ组PASMCs的增殖指数(24.67±3.19)则显著低于3个对照组(P<0.01),而AdCMVLacZ+ATⅡ组(59.53±3.26)和ATⅡ组比较无显著性差异。结论:反义AT1R通过抑制AT1R的表达而抑制PASMCs增殖。
Objective To investigate the influence of human angiotensin Ⅱ （AT Ⅱ ） type 1 receptor （AT1R） antisense cDNA （ahAT1） on proliferation of cultured human pulmonary artery smooth muscle cells （PASMCs）. Methods Two recombinant adenoviral vectors, AdCMVahAT1 containing full length antisense cDNA targeting to human AT1R mRNA, and AdCMVLacZ containing LacZ, were constructed by orientation clone and homologous recombination. The PASMCs was divided into 4 groups （ DMEM, AT Ⅱ , AdCMVLacZ ＋ AT Ⅱ and AdCMVahAT1 ＋ ATⅡ ） and corresponding intervention were given to different groups, respectively. AT1R expression was detected by RT - PCR and immunohistochemistry method. Proliferation index （PI） were determined by flow cytometry. Resutts After transfected 48 h, AT1R expression was significantly less in PASMCs transfected AdCMVahAT1 than that in group DMEM and in group AdCMVLacZ （ P 〈 0.01 ）. Stimulated by ATII for 48 h, in group AT Ⅱ the PI of PASMCs markedly increased （P 〈 0.01 Vs group DMEM）. However, in group AdCMVahAT1 ＋ATⅡ , PI of PASMCs markedly decreased （P 〈 0.01 Vs group ATⅡ and group DMEM, respectively）. There were no significant differences of PI between group AdCMVLacZ ＋ ATⅡ and group ATⅡ. Conclusion Human AT1 receptor antisense cDNA may decrease proliferation in human PASMCs by inhibiting AT1 R expression.
出处
《郧阳医学院学报》
2006年第5期257-260,276,F0002,共6页
Journal of Yunyang Medical College
基金
湖北省教育厅项目(B200524009)
关键词
反义AT1R
肺动脉
平滑肌细胞
增殖
Antisense AT1R
Pulmonary artery
Smooth muscle cell
Proliferation
