上海交通大学学报（医学版）

›› 2009, Vol. 29 ›› Issue (8): 935-.

• 论著（基础研究） • 上一篇    下一篇

反义Smad3阻断TGF-β1信号转导对血管平滑肌细胞增殖能力的影响

程智慧, 蔡文玮, 陆 平, 马绍骏, 陈 谊, 盛 净   

  1. 上海交通大学 医学院第九人民医院老年病科, 上海 200011
  • 出版日期:2009-08-25 发布日期:2009-09-27
  • 通讯作者: 盛净, 电子信箱: SHENGJING60@163.com。
  • 作者简介:程智慧(1981—), 女, 硕士生；电子信箱: zhihui9248527@sohu.com。

Effects of signal transduction interruption of transforming growth factor-β1 by anti-Smad3 on proliferation of vascular smooth muscle cells

CHENG Zhi-hui, CAI Wen-wei, LU Ping, MA Shao-jun, CHEN Yi, SHENG Jing   

  1. Department of Geriatrics, The Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200011, China
  • Online:2009-08-25 Published:2009-09-27

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摘要：

目的 观察反义Smad3腺病毒载体转染增生型血管平滑肌细胞(VSMC)后对其增殖能力的影响。 方法 构建大鼠颈总动脉急性球囊损伤模型。取损伤处血管组织，组织块贴壁法培养增生型VSMC，反义Smad3腺病毒载体转染（实验组）。以空载病毒作为阴性对照组，以无病毒载体作为空白对照组。抽提转染后细胞总RNA，RT-PCR检测转染后各组Smad3 mRNA表达；MTT比色法检测转染后细胞增殖能力的变化。 结果 与阴性对照组和空白对照组比较，转染后实验组Smad3 mRNA表达明显减少；转染后第2、3、5天，实验组细胞增殖能力明显降低(P<0.05)，转染后第7天各组细胞增殖能力差异无统计学意义（P>0.05）。 结论 反义Smad3腺病毒载体转染对增生型VSMC具有增殖抑制作用，为通过基因修饰阻断TGF-β1信号转导从而预防增生性血管病变奠定了基础。

关键词: 转化生长因子β1, 反义Smad3, 腺病毒, 血管平滑肌细胞, 增殖

Abstract:

Objective To observe the effects of anti-Smad3 adenovirus vector transfection on proliferation of proliferative vascular smooth muscle cells (VSMCs). Methods Rat models of acute balloon injury of common carotid artery  were established. The injured vascular tissues were obtained, proliferative VSMCs were cultivated by tissue explants adherent method, and anti-Smad3 adenovirus vector transfection was performed (experiment group). Virus without anti-Smad was served as negative control, and that without adenovirus vector was served as blank control. Total RNA of transfected cells was extracted, and the expression of Smad3 mRNA after transfection in each group was detected by RT-PCR. The changes in cell proliferation after transfection was determined by MTT. Results The expression of Smad3 mRNA in experiment group was significantly lower than that in negative control group and blank control group. Cell proliferation of experiment group was significantly decreased on day 2, 3 and 5 after transfection (P<0.05), while there was no significant difference in cell proliferation among each group on day 7 after transfection（P>0.05）. Conclusion Anti-Smad3 adenovirus vector transfection may inhibit the proliferation of proliferative VSMCs, which lays a foundation for the signal transduction interruption of TGF-β1 by gene modification in the prevention of proliferative vascular diseases.

Key words: transforming growth factor-β1, anti-Smad3, adenovirus, vascular smooth muscle cell, proliferation

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