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为探讨腺病毒介导反义人AT1R转染对培养的人血管平滑肌细胞 (VSMCs)增殖和凋亡的影响 ,用重组反义人AT1R的复制缺陷型腺病毒 (Ad CMV ahAT1)转染体外培养的人VSMCs,用RT PCR和免疫组化法检测AT1R的表达 ,用流式细胞仪检测VSMCs的增殖指数和凋亡率 ,绘制DNA合成直方图。结果显示 :与对照组比较 ,转染Ad CMV ahAT1后 4 8h的VSMCs,AT1RmRNA低 5 0 % ,AT1R蛋白呈低表达 (P <0 0 1) ;给予AngⅡ刺激的VSMCs的增殖指数明显增加 (与DMEM组比 ,P <0 0 1) ,转染Ad CMV ahAT1组的增殖指数则显著降低 (与DMEM组比P <0 0 5 ,与AngⅡ对照组比P <0 0 1)。Ad CMV ahAT1组在二倍体峰之前出现凋亡峰 ,各时相点细胞凋亡率分别为 18%(6h)、2 0 % (12h)、2 8% (2 4h)、2 6 % (4 8h)、3% (72h)和 1% (96h) ,而在其它各对照组未出现凋亡峰。这些提示腺病毒介导的反义人AT1R转染 ,通过抑制AT1R的表达 ,明显抑制人VSMCs的增殖和AngⅡ刺激的人VSMCs增殖 ,并诱导VSMCs凋亡。
In order to investigate the effect of adenovirus-mediated antisense AT1R transfection on the proliferation and apoptosis of cultured human vascular smooth muscle cells (VSMCs), the recombinant adenoviruses were transfected with Ad CMV ahAT1 recombinant antisense human AT1R Of human VSMCs. The expression of AT1R was detected by RT PCR and immunohistochemistry. The proliferation index and apoptosis rate of VSMCs were detected by flow cytometry, and the DNA synthesis histogram was drawn. The results showed that compared with the control group, VSMCs transfected with AdCMV ahAT1 had a 50% lower AT1R mRNA expression and a lower AT1R protein expression (P <0.01); the proliferation index of VSMCs stimulated with AngⅡ increased significantly ( DMEM group, P <0.01). The proliferation index of AdATV-ahAT1 group was significantly lower than that of DMEM group (P <0.05 compared with DMEM group, P <0.01). The apoptotic peak appeared before diploid peak in Ad CMV ahAT1 group, and the apoptotic rate was 18% (6h), 20% (12h), 28% (24 h), 26% 48h), 3% (72h) and 1% (96h), while no apoptotic peak appeared in other control groups. These suggest that adenovirus-mediated antisense human AT1R transfection, by inhibiting the expression of AT1R, significantly inhibited the proliferation of human VSMCs and the proliferation of human VSMCs stimulated by AngII, and induced the apoptosis of VSMCs.