腺病毒介导的rHSG-1基因转染对自发性高血压大鼠血管平滑肌细胞增殖的影响

Adenovirus-mediated gene transfer of rHSG-1 inhibits proliferation of vascular smooth muscle cells from spontaneously hypertensive rats

目的 :应用腺病毒介导的转基因方式转染大鼠增殖抑制基因 (rHSG 1) ,以观察rHSG 1对自发性高血压大鼠 (SHR)主动脉血管平滑肌细胞 (VSMCs)增殖的影响。方法 :体外培养大鼠主动脉血管平滑肌细胞并转染含有rHSG 1基因的重组复制缺陷型腺病毒载体 (Ad5rHSG 1) ,采用细胞计数、MTT和3 H thymidine参入法 ,观察rHSG 1对细胞增殖的影响 ;采用流式细胞仪分析rHSG 1对细胞周期的影响 ;应用WesternBlot方法检测rHSG 1对细胞周期调控蛋白p2 7Kip1、p2 1Cip1和增殖细胞核抗原 (proliferatingcellnuclearangitin ,PCNA)的作用。结果 :Ad5rHSG 1转染培养的SHRVSMCs后 ,能明显抑制细胞增殖 ,与对照组相比抑制率为 4 0 % (P <0 .0 1) ;细胞周期阻滞于G0 /G1期 ,周期调控蛋白p2 7Kip1、p2 1Cip1表达升高 ,PCNA表达降低。结论 :rHSG 1基因过表达可以抑制培养的自发性高血压大鼠血管平滑肌细胞增殖 ,并可能是通过参与细胞周期调节发挥作用。

Objective: To investigate the effect of rat hyperplasia suppressor gene-1(rHSG-1) on the proliferation of aortic vascular smooth muscle cells(VSMCs) from spontaneously hypertensive rats(SHR). Methods:VSMCs were transfected with an adenoviral vector expressing rHSG-1(Ad5rHSG-1). The effect of rHSG-1 on the proliferation of VSMCs was investigated by cell counting, MTT assay and 3H-thymidine incorporation. We also analyzed the cell-cycle using flow cytometry and detected the expression of p27 Kip1 and p21 Cip1 by Western Blot. Results:The proliferation of VSMCs infected with Ad5rHSG-1 was inhibited,a 40% reduction compared with the control group(P<0.01).The cell cycle was arrested in G 0/G 1 phase and the expression of p27 Kip1 and p21 Cip1 was upregulated after the VSMCs transfection with Ad5rHSG-1. Conclusion: Adenovirus-mediated gene transfer of rHSG-1 inhibits the cell cycle progression and thus the proliferation of VSMCs from SHR.