基础医学与临床 ›› 2014, Vol. 34 ›› Issue (7): 882-885.

• 研究论文 • 上一篇    下一篇

RAS信号途径参与干扰素-α抑制大鼠血管平滑肌细胞增殖

龙向淑1,吴强2,宋方2,黄晶3,张林军1   

  1. 1. 贵州省人民医院
    2. 贵州省心血管病研究所,贵州省人民医院
    3. 贵州省人民医院心血管内科
  • 收稿日期:2013-11-25 修回日期:2014-01-11 出版日期:2014-07-05 发布日期:2014-06-24
  • 通讯作者: 吴强 E-mail:waqqaa@yahoo.com.cn
  • 基金资助:
    贵州省优秀科技教育人才省长专项资金项目;家族性高胆固醇血症相关功能突变的组学筛查及基因敲除小鼠模型的建立

RAS signal pathway participation in interferon-α inhibition of proliferation of vascular smooth muscle cells in rats

  • Received:2013-11-25 Revised:2014-01-11 Online:2014-07-05 Published:2014-06-24

摘要: 目的 探讨RAS信号途径在干扰素-α(IFN-α)抑制大鼠血管平滑肌细胞(VSMCs)增殖中的作用。方法 应用转染IFI204 siRNA和/或IFN-α瞬时干预体外培养的大鼠VSMCs,以非特异性siRNA转染组为对照组,用MTT法测定细胞活力,流式细胞仪分析细胞周期,RT-PCR 法和Western blot分别检测P204 mRNA、P204和Ras蛋白的表达及RAF与ERK的磷酸化水平。结果 与对照组比较,IFN-α组P204 mRNA和蛋白表达上调(P<0.01),细胞活力和细胞周期G1/S转换下调(P<0.01),伴RAS蛋白表达减少(P<0.01),RAF及ERK磷酸化水平下降(P<0.01);转染IFI204 siRNA后再加IFN-α干预,P204 mRNA及蛋白表达下调(P<0.01),而G0/G1期细胞增多,S期细胞减少(P<0.01),且RAS蛋白表达和RAF及ERK磷酸化水平亦下调(P<0.01)。结论 RAS信号途径参与IFN-α抑制大鼠VSMCs增殖。

关键词: 干扰素, 血管平滑肌细胞, 增殖, 干扰素诱导蛋白,P204, RAS信号途径

Abstract: Objective To study the action of RAS signaling pathway in interferon alpha (IFN-α) inhibition proliferation of vascular smooth muscle cells (VSMCs) in rats. Methods Cultured VSMCs were treated with transfection IFI204 siRNA and/or IFN-αin vitro instantaneously. Nonspecific siRNA transfection group was as control group. The cell vitality was detected by MTT method, and the cell cycle was analyzed by flow cytometry. The expression of P204 mRNA was determined by semiquantitative RT-PCR. P204, RAS protein and the phosphorylation levels of RAF and ERK was analyzed by Western blotting. Results Compared with control group, the expression of P204 mRNA and protein up-regulated(P<0.01), the cell vitality and the cell cycle of G1/S transition down-regulated(P<0.01). In the process of the above, the expression of RAS protein decreased(P<0.01) and the phosphorylation levels of RAF and ERK droped(P<0.01); When intervention with IFN-α after transfection IFI204 siRNA, down-regulated the expression of P204 mRNA and protein(P<0.01), and cells of G0/G1 stage increased and S stage decreased(P<0.01), but the expression of RAS protein and the phosphorylation levels of RAF and ERK did not increase correspondingly(P<0.01). Conclusion RAS signal pathway participation in interferon-α inhibition of proliferation of vascular smooth muscle cells in rats.

Key words: Interferon, Vascular smooth muscle cells, Proliferation, Interferon-inducible protein, P204, RAS signaling pathway

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