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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。
AIM: To investigate the effect of adenovirus-mediated antisense AT1R cDNA transfection on the proliferation of human pulmonary artery smooth muscle cells (PASMCs). Methods: Recombinant antisense human AT1R adenovirus (AdCMVahAT1) and control vector AdCMVLacZ expressing β-galactosidase were constructed. The cultured PASMCs were divided into DMEM group, ATⅡ group, AdCMVLacZ + ATⅡ group and AdCMVahAT1 + ATⅡ group Corresponding intervention factors, the expression of AT1R was detected by RT-PCR and immunohistochemistry. The proliferation index of PASMCs in each group was detected by flow cytometry. Results: At 48 h after transfection, the expression of AT1R protein in AdCMVahAT1 group was significantly lower than that in AdCMVLacZ group and DMEM group. The proliferation index of PASMCs in ATⅡ group (59.69 ± 3.46) was higher than that in DMEM group (50.25 ± 1.34, P <0.01) at 48 h after ATⅡ stimulation. The proliferation index of PASMCs stimulated with ATⅡ for 48 h in AdCMVahAT1 + ATⅡ group (24.67 ± 3.19) was significantly lower than that of the three control groups (P <0.01), but there was no significant difference between AdCMVLacZ + ATⅡ group (59.53 ± 3.26) and ATⅡ group. Conclusion: Antisense AT1R inhibits the proliferation of PASMCs by inhibiting the expression of AT1R.