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Ski在全反式维甲酸抑制TGF-β1诱导的系膜细胞增殖中的作用

苏宝凤1,王晓花1,张莉1,于莹1,陈晓岚2   

  1. 1. 南京市第一医院
    2. 南通大学附属医院肾内科
  • 收稿日期:2012-08-17 修回日期:2012-11-26 出版日期:2013-04-15 发布日期:2013-04-15
  • 通讯作者: 苏宝凤

The effects of all-trans retinoic acid (ATRA) on the expression of Ski in Cultured rat mesangial cells treated by TGF-β1

  • Received:2012-08-17 Revised:2012-11-26 Published:2013-04-15 Online:2013-04-15
  • Contact: SU Bao feng

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摘要：

【摘要】目的 探讨全反式维甲酸（ATRA）对转化生长因子(TGF)-β1诱导的大鼠系膜细胞增殖及Ski表达的影响。方法 不同浓度ATRA预处理大鼠HBZY-1系膜细胞（为各剂量ATRA组）24 h后再加TGF-β1 (10 μg/L)培养24 h。 CCK-8法检测细胞增殖情况；Real-time PCR法检测Ski mRNA的表达；Western blot检测Ski蛋白的表达；激光共聚焦荧光显微镜检测Ski蛋白的亚细胞定位。并与正常对照组及TGF-β1组比较。结果 与正常对照组相比，TGF-β1组系膜细胞增殖明显，且Ski mRNA及蛋白表达升高(P < 0.05)；与TGF-β1 组相比，ATRA能够呈剂量依赖性地抑制 TGF-β1的促增殖作用，ATRA 10 μmol/L组Ski mRNA及蛋白表达量明显增高。ATRA组Ski蛋白主要定位于大鼠系膜细胞核，其细胞核荧光信号强度较TGF-β1组明显增强，细胞浆中荧光信号强度较TGF-β1组减弱。结论 ATRA可通过上调大鼠系膜细胞Ski表达，抑制其向核外转位，从而抑制TGF-β1诱导的系膜细胞增殖。

关键词: 大鼠系膜细胞, 全反式维甲酸, 转化生长因子-β1, Ski

Abstract:

[Abstract] Objective  To investigate the effects of all-trans retinoic acid (ATRA) on the expression of Ski in cul tured rat mesangial cells treated by transforming growth factor (TGF-β1). Methods  The mesangial cells were pre-treated with different concentrations of ATRA for 24 h, and then stimulated with TGF-β1 (10 μg/L) for 24 h. The cell proliferation was detected by CCK-8 method. The Ski mRNA expression was determined by real time-PCR. The protein expression of Ski was determined by Western blot analyses. The localization of Ski was observed by confocal fluorescence microscopy. Results  Compared with the control group, TGF-β1 significantly enhanced the proliferation and the expression of Ski mRNA and protein in rat mesangial cells (P < 0.05). ATRA significantly inhibited mesangial cell proliferation induced by TGF-β1 in a dose-dependent manner. In ATRA groups, Ski mRNA and protein were increased significantly. Additionally, Ski protein was mainly localized in nucleus, and fluorescence intensity was significantly increased in nucleus, while decreased in cyto? plasm in ATRA groups. Conclusion  ATRA could inhibit TGF-β1 induced mesangial cell proliferation through up-regulat? ing the expression of Ski and inhibiting it translocation from the nucleus to the cytoplasm.

Key words: Rat Mesangial cells, All-trans retinoic acid, Transforming growth factor –β1, Ski